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ივ. ჯავახიშვილის სახელობის თბილისის სახელმწიფო

უნივერსიტეტი

ხელნაწერის უფლებით

გოჩა ომარის ძე სირბილაძე

აპოპტოზის როლი და მისი მოდულაცია იმუნური პასუხით

განპირობებული ღვიძლის მსუბუქი დაზიანების დროს

14.00.36_ალერგოლოგია და იმუნოლოგია

დ ი ს ე რ ტ ა ც ი ა

ბიოლოგიის მეცნიერებათა კანდიდატის

სამეცნიერო ხარისხის მოსაპოვებლად

სამეცნიერო ხელმძღვანელი

მედიცინის მეცნიერებათა დოქტორი

თინათინ ილარიონის ასული ჩიქოვანი

თბილისი

2006

2

Sinaarsi

Semoklebebis nusxa ...................................................................................................................4

Sesavali.................................................................................................................................................5

I Tavi - literaturis mimoxilva.........................................................................12

1.1 imunuri pasuxiT ganpirobebuli RviZlis dazianebis meqanizmebi .........12

1.2 apoptozis roli RviZlis normalur funqcionirebaSi ................................277

1.3 kaspazebis roli apoptozSi ....................................................................................................... 36

1.4 apoptozis modulacia Terapiuli saSualebebiT ............................................... 400

II Tavi - sakvlevi masala da meTodebi......................................................... 45

2.1 RviZlis imunuri dazianebis eqsperimentuli modeli....................................... 45

2.2 histomorfologiuri meTodi .................................................................................................... 45

2.3 RviZlSi apoptozis Sefaseba imunohistoqimiuri (TUNEL)LmeTodiT ...45

2.4Aazotis oqsidisa da glutaTion reduqtazas atiurobis gansazRvra

........................................................................................................................................................................................46

2.5 hepatocitebis gamoyofa da maTi mitoqondriuli aqtiurobis

Seswavla tetrazoliumze dafuZnebuli kolorimetriuli MTT testiT

........................................................................................................................................................................................47

2.6 hepatocitebis kaspazebis aqtiurobis gansazRvra................................................48

2.7 ALT-s gansazRvris speqtrometriuli meTodi .......................................................... 48

2.8 masalis statistikuri damuSaveba........................................................................................49

III Tavi - sakuTari gamokvlevis Sedegebi .................................................. 50

3.1 plaferoni lb-s zegavlena Con-A inducirebuli msubuqi hepatitis

dros sisxlSi alaninaminotransferazas Semcvelobasa da RviZlSi

ganviTarebul morfologiur cvlilebebze........................................................................ 50

3.2 Con-A inducirebuli msubuqi hepatitis dros RviZlsa da sisxlSi

mimdinare oqsidaciuri procesebis maCveneblebis modulacia

imunomodulatorebiT ............................................................................................................................... 59

3.3 Con-A inducirebuli msubuqi hepatitis dros RviZlSi kaspazebis

aqtiuroba da maTi modulacia imunomodulatorebiT .............................................64

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3.4 eqsperimentuli msubuqi hepatitis dros hepatocitebis apoptozuri

cvlilebis Seswavla imunohistoqimiuri meTodiT........................................................67

IV Tavi – sakuTari Sedegebis ganxilva........................................................ 67

daskvnebi .............................................................................................................................................................. 833

praqtikuli rekomendaciebi.................................................................................................................84

literaturis sia........................................................................................................................................... 85

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Semoklebebis nusxa

dnm/rnm – dezoqsi/ribonukleinis mJava

kaspaza – apoptosis mwarmoebeli cisteinuri proteaza

ALT – alaninaminotransferaza

Apaf-1 – adapteri proteinis apoptozuri proteaz-gamaaqtiurebeli faqtori-1

ATP - adenozintrifosfati

Bcl-2 – B-ujreduli leikemiis proteini

Bid, Bak, Bax – Bcl-2 ojaxis proapoptozuri wevrebi

CD – diferencirebis klasteri

Con-A – konkanavalin-A (bunebrivi mitogeni)

CTL – citotoqsiuri A limfocitebi (bunebrivi mitogeni)

GalN – d-galaqtozamini

GSH – glutaTioni

GVHD - transplantanti maspinZlis winaaRmdeg” daavadeba

FADD - FAS-asociirebuli sikvdilis domeni

FAS – simsivnis nekrozis gaqtoris ojaxSi Semavali II tipis transmembranuli

proteini

HBV - hepatitis B virusi

HCV - hepatitis C virusi

ICAM – ujredSorisi adheziuri molekula

IFN – interferoni

IL – interleikini

HbsAg - hepatitis B virusis zedapiruli antigeni

iNOS - inducibeluri azotis oqsidis sinTetaza

LPS – lipopolisaqaridi (endotoqsini)

MHC - histoSemTavseblobis mTavari kompleqsi

NF-kB - birTvis transkripciuli faqtori kappa-B

NK - bunebrivi kilerebi /imunuri ujredebi/

NO - azotis oqsidi

NOS - azotis oqsidis sinTetaza

RIP – receptorTan urTierTmoqmedi proteini

ROS – reaqtiuli Jangbadis saxeobebi

TNF – citokin simsivnis nekrozis faqtoris jgufis warmomadgenelbi

TGF - simsivnis zrdis faqtori

VCAM – sisxlZarRvovani adheziuri molekula

5

Sesavali

problemis aqtualoba

medicinis ganviTarebis mniSvnelovani tempebis miuxedavad,

XXI saukunis dasawyisSi virusuli hepatitebi kvlav mZime da

globalur problemad rCeba. jandacvis msoflio organizaciis

monacemebiT virusuli hepatitiT daavadebulTa raodenoba

ganuxrelad matulobs da miliardze mets Seadgens. ufro metic,

sxvadasxva saxis hepatitiT dedamiwis mosaxleobis TiTqmis mesamedi

aris an odesme iyo inficirebuli. msoflioSi yovelwliurad am

daavadebiT an misi garTulebiT 2 mln-mde adamiani iRupeba.

saqarTveloSi 2004 wels virusuli hepatitebis (B da C) 1000-

mde axali SemTxveva dafiqsirda. aRsaniSnavia, rom B hepatitis

SemTxvevaSi mwvave mimdinareoba aRiniSneboda daavadebulTa 64%-Si,

xolo qronikuli – 35%-Si, maSin roca C hepatitis dros faqtiurad

naxevarze meti (daaxloebiT 55%) qronikuli iyo. ufro metic,

saqarTvelos mosaxleobis 5-7% hepatitis sxvadasxva virusis

matarebelia.

B da C hepatitis virusebiT dainficireba organizms ori

mTavari saSiSroebis winaSe ayenebs: erTi mxriv, daavadebis

fulminanturi mimdinareobis SemTxvevaSi (RviZlis koma, elviseburi

hepatiti) maRali letaluri gamosavali (90% letalurad

mTavrdeba), meores mxriv ki, procesis qronizacia, rac sabolood

RviZlis cirozis an hepatoceluluri karcinomis ganviTarebiT

mTavrdeba. hepatitis elviseburad mimdinareobas adgili aqvs B da D

hepatitebis virusiT erTdroulad dainficirebis 80%-Si.

msoflios wamyvani hepatologiuri centrebis monacemebiT,

HBV infeqciis fulminanturi mimdinareobis saSiSroeba

inficirebulTa 80%-Si SeiniSneba (Grob et al., 1998). msoflio

jandacvis organizaciis monacemebiT, daaxloebiT 2 miliard

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adamians gadatanili aqvs B hepatiti, xolo 350 milionze meti HBV

mtarebelia, qronikul mtarebelTa 25%-Si ki daavadeba RviZlis

kibos ganviTarebiT mTavrdeba (Alter, 1995). msoflioSi RviZlis

kiboTi yovelwliurad 1 milionze meti adamiani iRupeba. qronikuli

HBV infeqciis persistentobis erT-erTi mizezi isaa, rom

inficirdeba hepatocitebis 100% da ar xdeba maTi mocileba.

mwvave virusuli hepatitis gadatanis Semdeg

gamojanmrTelebis procesi didxans mimdinareobs. es gansakuTrebiT

exeba C hepatits, romelic yvelaze rTuli mimdinareobiT

xasiaTdeba. msoflio jandacvis organizaciis monacemebiT, C

hepatitiT msoflioSi 300 milioni adamiania inficirebuli,

romelTagan 130 milioni am virusis qronikuli mtarebelia. C

hepatiti SemTxvevaTa 70-80%-Si ganicdis qronizacias da amaTgan 50-

%-s cirozi uviTardeba. ufro metic, zogjer qronikuli hepatitis

diagnostireba ar xdeba da daavadeba mxolod cirozis stadiaze

gamovlindeba. sakmaod xSirad C hepatitis mqone pacientebs

axasiaTebT “cru gamojanmrTeleba”, roca sisxlis bioqimiuri

analizebi normaSi modis (Scheuer et al, 1990). es fenomeni SeiZleba

gagrZeldes ramdenime Tvidan ramdenime wlamdec ki. amitom arc Tu

usafuZvlod mas simZimis mixedviT Sidss adareben. yovelive zemoT

Tqmulidan gamomdinare, virusuli hepatitebis mkurnalobas Zalian

didi mniSvneloba aqvs.

aracitopaTiuri virusebiTa (HBV, HCV) da autoimunuri

geneziT gamowveuli hepatitebis dros RviZlis dazianebis

meqanizmebi sabolood garkveuli ar aris, Tumca masSi T ujreduli

imunuri pasuxis ganmsazRvreli roli dReisaTvis dasabuTebulia

(Ando et al., 1997). adekvaturi imunuri pasuxi praqtikulad

gamoricxavs qronikuli hepatitis ganviTarebis Sanss. susti

imunuri pasuxis SemTxvevaSi daavadebis mwvave faza msubuqad,

xSirad usimptomod mimdinareobs, rac SesaZloa procesis Semdgom

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qronizacias da hepatoceluluri karcinomis an cirozis

ganviTarebas ganapirobebdes.

RviZlis funqcionirebis SenarCunebaSi gadamwyveti

mniSvneloba apoptozis saSualebiT virusiT inficirebuli

ujredebis swraf da droul mocilebas eniWeba. janmrTel

organizmSi mkvdari ujredebi swrafad Cainacvleba mitozis gziT

warmoSobili axali ujredebiT, anu organoebSi ujreduli

homeostazi narCundeba, maSin roca mwvave dazianebis dros Warbi

da/an xangrZlivi apoptozi RviZlis funqciisa da reperfuziis

moSlis mizezi xdeba. Warbi apoptozi aRiniSneba mwvave virusuli,

alkoholuri hepatitebis, RviZlis qolestazuri daavadebis,

vilsonis daavadebisa da “transplantanti maspinZlis winaaRmdeg”

daavadebisTvis (GVHD). meores mxriv, virus-inficirebuli

hepatocitebis apoptozis deficiti mutirebuli genebis matarebeli

ujredebis mocilebis moSlasa da ujredebis usazRvro

gamravlebas iwvevs, rac gamudmebuli anTebis pirobebSi RviZlisa

da naRvlsadeni sistemis simsivneebis ganviTarebas uwyobs xels.

aqedan gamomdinare, sxvadasxva paTologiis dros RviZlis qsovilSi

apoptozis mainhibirebeli Tu mastimulirebeli preparatebi

potenciurad Zlieri Terapiuli saSualebebi SeiZleba gaxdes.

virusuli hepatitebis paTogenezidan gamomdinare, zogierTi

avtori virusuli hepatitebis samkurnalod upiratesobas

imunotropul da antioqsidaciur praparatebs aniWebs (Zein et al., 1998;

Schwarz, 1996; d.metreveli, 1999; d. pavliaSvili, 1999). aRniSnuli

faqtis gaTvaliswinebiT sainteresoa iseTi imunomamodulirebeli

Tvisebis mqone preparatebis Seswavla, rogoricaa plaferoni lb.

plaferoni lb Seqmnilia samedicino bioteqnologiis

institutSi profesor vladimer baxutaSvilis mier. mas rogorc

imunomamodulirebeli, ise anTebis sawinaaRmdego, antiiSemiuri da

antioqsidaciuri Tvisebebi gaaCnia (T. Ciqovani, 1997; d. WavWaniZe,

1999; r. ruxaZe, 1999; m. gongaZe, 2004). garda amisa, dadginda, rom

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plaferoni lb-s gamoyeneba RviZlis sxvadasxva paTologiis dros

RviZls metaboluri da morfologiuri cvlilebebis

ganviTarebisgan icavs (m. faSalaSvili, 1995; d.pavliaSvili, 1999;

Rukhadze et al., 1998; Chikovani et al., 1999). ufro metic, Seswavlilia

plaferoni lb-s antiapoptozuri Tvisebac (WoWua da sxv., 2006).

preparatis hepatoproteqtoruli moqmedebis zusti meqanizmi ki

dRemde ucnobia.

yovelive zemoTTqmulidan gamomdinare, mizanSewonilad

CavTvaleT, Segveswavla apoptozis roli Con-A inducirebuli

msubuqi hepatitis eqsperimentul modelSi da masze

imunomamodulirebeli preparatebis gavlena.

yovelive zemoT Tqmulidan gamomdinare, mizanSewonilad

CavTvaleT, Segveswavla apoptozis roli Con-A inducirebuli

msubuqi hepatitis eqsperimentul modelSi da masze

imunomodulatorebis moqmedebis meqanizmi.

kvlevis mizani da amocanebi

kvlevis mizans eqsperimentuli msubuqi hepatitis dros

RviZlSi ganviTarebuli apoptozuri cvlilebebisa da masze

imunomodulatorebis gavlenis Seswavla warmoadgenda.

aRniSnuli miznis misaRwevad Semdegi konkretuli amocanebis

gadawyveta daisaxa:

1. Con-A-inducirebuli msubuqi hepatitis eqsperimentuli

modelis SemuSaveba (konkanavalin A-s dozis SerCeva

transaminazebis donis cvlilebiT);

2. imunomodulatorebis (plaferoni lb, deqsametazoni) gavlenis

Seswavla Con-A inducirebuli msubuqi hepatitis dros

plazmaSi transaminazebis aqtivobaze;

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3. Con-A inducirebuli msubuqi hepatitis dros kaspazebis (3, 8)

aqtiurobisa da maTze imunomodulatorebis gavlenis

Seswavla;

4. imunomodulatorebis gavlenis Sefaseba Con-A inducirebuli

msubuqi hepatitis dros RviZlSi ganviTarebul histologiur

cvlilebebze;

5. imunomodulatorebis gavlenis Seswavla Con-A inducirebuli

msubuqi hepatitis dros sisxlSi ganviTarebuli oqsidaciuri

procesebis maCveneblebze (oqsidanturi da antioqsidanturi

sistemis Sefaseba).

kvlevis mecnieruli siaxle

winamdebare naSromSi pirvelad dadginda, rom msubuqi

formis hepatitis dros RviZlSi Zlierdeba kaspaza 3 da 8-T

warmoebuli apoptozi.

dadginda, rom imunomodulatorebis gamoyeneba Con-A-

inducirebuli msubuqi hepatitis dros RviZls icavs paTologiuri

cvlilebebis ganviTarebisagan. amasTanave, plaferoni lb RviZlis

qsovilSi ganviTarebuli cvlilebebis swraf aRdgenas

uzrunvelyofs.

pirvelad iqna naCvenebi, rom msubuqi hepatitis dros

Pplaferoni lb zrdis kaspazebis 3, 8 aqtiurobas da amiT xels

uwyobs dazianebuli ujredebis mocilebas apoptozis gziT.

dazianebuli ujredebis moSoreba ki, Tavis mxriv, aRkveTs

daavadebis qronizaciis SesaZleblobas.

naSromis praqtikuli Rirebuleba

winamdebare naSromis Sedegebma gamoavlina plaferoni lb-s

proapoptozuri Tviseba da cxadyo misi moqmedebis zogierTi

molekuluri meqanizmi, rac safuZvlad daedeba mis gamoyenebas

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aramarto mwvave virusuli hepatitebis, aramed, RviZlis qronikuli

infeqciis dros.

dacvaze gamotanili ZiriTadi debulebebi

msubuqi hepatitis eqsperimentul modelSi plaferoni lb-s

gamoyeneba aCqarebs RviZlSi regeneraciuli procesebis

ganviTarebas.

plaferoni lb-s gamoyeneba Con-A-inducirebuli msubuqi

hepatitis eqsperimentul modelSi zrdis hepatocitebSi

kaspazebis (3 da 8) aqtiurobas RviZlis normaluri morfo-

funqciuri mdgomareobis aRdgenis periodSi.

plaferoni lb eqsperimentis sxvadasxva vadaze sisxlSi

azotis oqsidis Semcvelobaze gansxvavebulad moqmedebs.

eqsperimentuli msubuqi hepatitis dros imunomodulatorebis

gamoyeneba sisxlSi glutaTionreduqtazas dones

aregulirebs.

naSromis aprobacia

disertacia aprobirebulia saqarTvelos mecnierebaTa

akademiis samedicino bioteqnologiis institutSi biomedicinisa da

bioteqnologiis ganyofilebebis gafarToebul sxdomaze 2006 wlis

3 maiss. disertacia rekomendirebulia sajaro dacvisaTvis.

disertaciis ZiriTadi debulebebi moxsenebulia

I saerTaSoriso konferencia _ “samedicino reabilitacia da

raitTerapia” (Tbilisi, 20-24 ivnisi, 2006).

disertaciis moculoba da struqtura

disertacia warmodgenilia nabeWdi teqstis 101 gverdze.

Seicavs Sesavals, literaturis mimoxilvas, masalisa da meTodikis

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aRweras, sakuTar gamokvlevebs, kvlevis Sedegebis analizs,

daskvnebs, praqtikul rekomendaciebs da gamoyenebuli literaturis

sias. naSromi gaformebulia 2 sqemiT, 1 cxriliT, 6 diagramiT da 13

suraTiT. literaturis sia warmodgenilia 185 dasaxelebiT (maT

Soris 10 qarTulenovani da 175 inglisurenovani wyaroTi).

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I Tavi - literaturis mimoxilva 1.1 imunuri pasuxiT ganpirobebuli RviZlis dazianebis

meqanizmebi

dReisaTvis hepatitebis gamomwvevi ramdenime virusia cnobili.

hepatitis virusi pirdapiri citopaTiurobiT ar gamoirCeva (Lok et al.,

2001). aqedan gamomdinare, hepatocitebis lizisi organizmis imunuri

pasuxis siZlierezea damokidebuli (Dudley et al., 1972). autoimunuri

genezisa da aracitopaTiuri virusebiT (HBV, HCV) gamowveuli

hepatitebis dros RviZlis dazianebas T ujreduli reaqcia

ganapirobebs. virusuli etiologiis RviZlis dazianebis dros

ujred-damokidebuli imunuri pasuxi mniSvnelovan rols TamaSobs

hepatocelularuli nekrozisa da imunopaTogenezuri meqanizmebis

ganviTarebaSi. cnobilia, rom RviZlis mwvave dazianebas

paTologiuri apoptozi ganapirobebs (Gremion et al., 2004; Kountouras et

al., 2003). apoptozis defeqtma ki SeiZleba avTvisebiani

transformacia, virusuli latenturi mimdinareoba da autoimunuri

daavadebebi gamoiwvios (Solary et al., 1996).

virusuli hepatitebis dros RviZlis qsovilSi saerTo

histologiuri cvlilebebi viTardeba. kerZod, damaxasiaTebeli

niSania RviZlis mwvave anTeba da qronizaciis SemTxvevaSi

fibrozuli gadagvareba. anTebiTi procesebis dros xSirad xdeba

RviZlSi limfocitebiT infiltracia, hepatocitebis masiuri

nekrozi, romelsac Tan erTvis leikocituri da histocituri

reaqcia (Adams et al., 1989).

13RviZlis dazianeba

mwvave qronikuli

citokinebi, oqsidaciuri stresi

hepatocitebi stelaturi ujredebi

nekrozi apoptozi fibrozi

RviZlis funqciuri ukmarisoba

sqema #1. apoptozis sqematuri roli RviZlis daavadebebis ganviTarebaSi

masiuri fulminanturi nekrozis dros mTeli acinusi

ziandeba. RviZli zomaSi mcirdeba. nekrozis farTo gavrcelebisas

adgili aqvs qsovilis nekrozs SemaerTebelqsovilovani xarixebis

SenarCunebiT. am nekrozs “mastoidurs” uwodeben. Sedegad,

sabolood SeiZleba cirozi ganviTardes. xSirad mas nawiburis

ganviTareba sdevs Tan.

hepatocelularuli dazianeba umetes SemTxvevaSi gamowveulia

ara uSualod damazianebeli agentis, aramed im anTebis ujredebis

mier, romlebic stresis qveS myofma hepatocitebma moizides (Ando et

al., 1997). RviZli kritikul organoa adamianis sxva simsivnebis drosac.

hepatocelularuli karcinoma msoflioSi erT-erTi yvelaze

gavrcelebuli mzardi sixSiris simsivnia. garda amisa, RviZli

metastazis gamo sxva simsivnebis umravlesobis samiznecaa.

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hepatocelularuli karcinoma metad axlos dgas qronikul

hepatitTan da cirozTan, rac regeneraciis dros zrdis kontrolis

meqanizmebSi momxdar cvlilebebze miuTiTebs. es ki, Tavis mxriv,

nawilobriv citokinebis moqmedebis Sedegia da RviZlis

karcinogenezSic SeiZleba hqondes adgili.

normalur hepatocitebs sxvadasxva citokinis (interleikin-1

alfas (IL-1), interleikin-1 betas (IL-1), simsivnis nekrozis

faqtoris (TNF), IL-6-is) receptorebi gaaCnia. garda amisa, RviZlis

ujredebis umetesobas, rogoricaa kupferis ujredebi, hepatocitebi,

da limfocitebi, citokinebis sinTezisa an maTze reagirebis unari

Seswevs. amdenad, RviZli citokinebis rogorc mTavar mwarmoebels,

ise maT gamanadgurebels warmoadgens. amis gamo, RviZlis daavadebis

mravali tipi citokinebis metabolizmis regulirebis moSlas

ukavSirdeba. RviZlis daavadebis mizezisda mixedviT, ramdenime

infeqciuri, arainfeqciuri da metaboluri agenti citokinebis

gamomuSavebis mTavar stimulebs SeiZleba warmoadgendes.

mwvave virusuli hepatitebis dros izrdeba IL-6, TNFα, IL-1 da

IL-2 koncentraciebi, romlebsac aqvT pro-anTebiTi da citotoqsiuri

efeqti (Bertoletti et al., 2003). qronikuli HBV-s dros IL-2 da TNFα-s

Semcveloba mcirdeba da izrdeba IL-1 da IL-2R done. qronikuli

HCV-s dros ki IL-2R izrdeba, xolo IL-2 da IFNγ mcirdeba.

hepatitebis dros gazrdili virusuli replikacia ROS-

damokidebuli birTvis transkrifciis faqtor kappa-B-s

gamonTavisuflebas iwvevs (Schwarz, 1996). mwvave virusuli

hepatitebis dros IL-1α, IL-1β, IL-6 da TNFα-s done izrdeba (Tsutsui et al.,

2003), dabalia lipiprotein(α)-s done (Geiss et al., 1996).

imunuri pasuxi virusis eliminaciaSi udides rols TamaSobs,

rac damtkicebuli iqna mwvave B da C hepatitebs Seswavlisas. mwvave

B hepatitis mimdinareoba HBV-s mimarT citotoqsiuri T-

limfocitebis (CTL) poliklonuri pasuxiT xasiaTdeba, rac ar

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SeiniSneba qronikuli B hepatitiT daavadebulebSi. dadgenilia, rom

daavadebis gamosavali HBV-specifiur CTL pasuxzea damokidebuli

(Rehermann, 1996).

mwvave B hepatitis dros HBV-s oligopeptidebi ukavSirdebian

maspinZlis ujredebis MHC I klasis molekulebs da

transportirdebian ujredis zedapirze. aseT ujredebs T-

limfocitebi aRmoaCenen da anadgureben (Kakimi et al., 2000). Tu ar

moxda virusis eliminacia, procesi qronikul xasiaTs iRebs. mwvave

B hepatitis keTilsaimedo mimdinareobis dros imunuri sistema

virusis eliminacias uzrunvelyofs manam, sanam ujredis genomSi

virusis genomis saboloo integracia moxdeba. HBV infeqciis

imunopaTogenuri meqanizmebis ganviTarebaSi erT-erTi mniSvnelovani

roli citokinebs ekuTvnis. isini imunuri sistemis modulacias

axdenen da pirdapiri antivirusuli moqmedeba gaaCniaT (Missale et al.,

1995). mwvave B hepatitis fulminanturi mimdinareobis dros

momatebulia IL-1, IL-2, IL-6 da TNFα (Webster et al., 2000; Missale et al., 1995).

normaSi hepatocitebi gamoyofen ujredSorisi adheziis molekula-1-

is (ICAM-1) Zalian mcire raodenobas, rac aRmoCenili iqna

imunopaTologiurad. ICAM-1-is Warbi eqspresia ujredebze RviZlis

anTebiTi dazianebis mauwyebelia, rasac xels uwyoben IL-1β, TNFα,

IFNγ da endotoqsini (Satoh et al., 1996). B hepatitis dros

inducirebuli periferialuri sisxlis mononukleuri ujredebis

sikvdili SeiZleba IL-12 heterodimeris da IL-10-is eqspresiis

anormalurobasTan iyos dakavSirebuli (Ji et al., 2001). garda amisa,

hepatitis B virusi zrdis iNOS-is transkrifcias ujredul

kulturebSi (Amaro et al., 1999). unda aRiniSnos, rom mwvave B

hepatitis dros periferiuli sisxlis mononukleuri ujredebis

spontanuri porliferaciis matebis fonze am ukanasknelTa pasuxi

mitogenur stimulaciaze daTrgunulia, rac limfocitebisa da

16

monocitebis funqciuri aqtivobis daqveiTebiT aris gamowveuli (d.

pavliaSvili 1999).

virusis persistencia SesaZloa damokidebuli iyos T

ujredebis specifiur defeqtTan, romelic HBV antigenis

gamocnobas uSlis xels (Zheng et al., 2004). rogorc zemoT aRvniSneT,

qronikuli hepatitebis dros ganviTarebuli imunuri pasuxi sustia.

Tu qronikuli B hepatitis dros imunuri pasuxi Zalian sustia,

maSin RviZlis dazianebac naklebia an saerTod ar aRiniSneba,

xolo virusi replikacias agrZelebs RviZlis normaluri funqciis

fonze. aseTi avadmyofebi “janmrTeli” matareblebi xdebian. amas

gansakuTrebuli mniSvneloba aqvs leikemiiT daavadebulTaTvis,

Tirkmlis ukmarisobis an gadanergili organos mqone pirTaTvis,

imunosupresuli mkurnalobis qveS myofi pacientebisTvis, SidsiT

daavadebulTaTvis, axalSobilebisTvis. maT RviZlSi sakmao

raodenobiT HBsAg-ia aRmoCenili, miuxedavad, imisa, rom ar

arsebobs hepatocelularuli nekrozi. Tu ujred-damokidebuli

imunuri pasuxi ufro gamoxatulia, maSin hepatoceluluri nekrozi

viTardeba, magram es pasuxi sakmarisi ar aris virusis

eliminaciisaTvis, sabolood ki qronikuli hepatiti viTardeba

(Dudley et al., 1972).

ase rom, B hepatitis gamosavali humoruli da ujreduli

imunitetis mdgomareobiT ganisazRvreba.

virusiT infiltrirebuli hepatocitebis lizisis

SeuZlebloba mravali meqanizmiT aixsneba. igi SeiZleba

dakavSirebuli iyos supresoruli T ujreduli funqciis

gaZlierebasTan, citotoqsiuri limfocitebis (kilerebis)

defeqtTan an ujredis membranaze antisxeulebis blokirebasTan.

qronikuli B hepatitis dros antigeniT gaaqtivebuli

leikocitebi citokinebs gamoimuSaveben da xdeba araspecifiuri

imunuri pasuxis gacema. aRniSnuli citokinebi moicavs

17

interferonsac. organizmi infeqcias interferonis produqciiT

pasuxobs, rac 2 efeqts iZleva: enzimebis (mag: 2’5’-

adeniladsinTataza) lokaluri stimulacia, romlis saSualebiTac

virusis dnm-is replikacia inhibirdeba, da imunuri pasuxis

stimulacia, romelic citotoqsiuri limfocitebis da NK

ujredebis aqtivaciiT xorcieldeba (Ratajczak, 1993). zogierT

SemTxvevaSi daqveiTebulia interferonis produqcia, rac

hepatocitebis membranaze adamianis I klasis leikocituri antigenis

eqspresias uSlis xels. aqtiuri HBV infeqciis dros RviZlSi

lokalurad producirdeba IFNγ, IL-1da TNFα. (Tsutsui et al., 2003).

SesaZloa, es anTebis araspecifiuri gamovlineba iyos. am dros

aRmoCenilia agreTve IL-2, IL-4, IL-5-is matriculi rnm-ebi. analizebma

cxadyo, rom IFNγ-s absoluturi done mimarTulia RviZlis mZime

anTebiTi procesis korelaciisken (Shindo et al., 1996).

qronikuli hepatitis dros gamomwvevi virusis mimarT susti

imunuri pasuxi viTardeba (Jeroen et al., 2005). aRsaniSnavia, rom

imunuri reaqciebi am dros hepatocitebis membranis

komponentebisaken aris mimarTuli. amasTanave, naCvenebia ujreduli

imunuri pasuxi hepatocitebis antigenebis mimarT. aRniSnuli

imuniteti gaaqtiurebuli limfocitebiT da mononukleuri

ujredebiT xorcieldeba. qronikuli hepatitebis dros

gaaqtivebuli ujredebis mier gamoTavisufldeba citokinebi,

romlebic inicireben wamyvan kaskadur reaqciebs, rac paTologiis

aRmofxvrisaken aris mimarTuli (Streetz et al., 2001). IL-1β hepatocitebis

Fas–damokidebuli apoptozis supresias axdens. es ki kaspaza 3-is

supresiiT xorcieldeba, romelic apoptozis kontrols axdens

(Takehara et al., 1999). IL-2 aaqtiurebs NK ujredebs, romlebsac

hepatocitebis apoptozi da nekrozi SeuZliaT gamoiwvion (Blom et al.,

1999).

18

1989 wels C hepatitis virusis identifikacia moxda.

statistikuri monacemebiT, msoflios donorTa 0.01-2%-is sisxli C

hepatitiT aris inficirebuli (Alter et al., 1995), xolo im pirebs,

romlebsac hemotransfuzia mravaljeradad utardebodaT,

SemTxvevaTa 10-15%-Si anti-HCV gamouvlindaT (Makris et al., 1990). HCV-

inficirebuli nemsiT Cxvletisas avaddeba 3-10% (Mitsui et al,. 1992).

aSS-Si narkomanTa 40% mwvave C hepatitiT aris daavadebuli

(saerTo Spricebis gamoyenebis gamo), xolo did qalaqebSi mcxovreb

narkomanTa 28%-s infeqciidan 2 wlis Semdeg sisxlSi anti-HCV

antisxeulebi aReniSneba. fiqroben, rom sqesobrivi gziT C

hepatitiT inficirebis albaToba umniSvneloa (Bresters et al., 1994).

agreTve naklebad aRiniSneba virusis vertikalurad gadacemis

SemTxvevebic. misi sixSire im SemTxvevaSi matulobs, roca dedis

sisxlSi HCV–is rnm aris momatebuli (Ohto et al., 1994).

aRsaniSnavia, rom HCV virusi cirkulirebs RviZlsa da

sisxlSi, xolo sxeulis sxva nawilebSi igi ar moipoveba. virusi

gamoirCeva Zlieri rezistentulobiT garemo pirobebisadmi da

daRvrili sisxlis gamSral laqebSi TveobiT cxovrobs (Liang et al.,

2000).

C hepatiti ufro xSirad usimptomod mimdinareobs da

daaxloebiT 70-80%-Si qronikul xasiaTs iZens. e.i. mwvave C hepatiti

inficirebulTa mxolod mcire nawilSi aRiniSneba.

C hepatitis gamomwvevi virusis organizmSi moxvedra

procesSi sxvadasxva imunuri stimulis CarTvas ganapirobebs (Lauer

et al., 2001). izrdeba IL-2, IL-6, IL-10 da IFNα done (Mihm et al., 1997),

matulobs alaninaminotransferazas Semcveloba, irRveva oqsidant-

antioqsidanturi sistema, rac lipidebis zeJanguri Jangvis

gaaqtivebasa (Paradis et al., 1997) da oqsidaciuri stresis ganviTarebas

uwyobs xels (Mutlu-Tarkoglu et al., 1997). Tavisufali radikalebis

momateba glutaTionis da malondialdehidis gaaqtiurebas iwvevs

19

(Farinati et al., 1996), yovelive zemoT Tqmuli ki xels uwyobs RviZlis

dazianebas da fibrozis ganviTarebas. HCV-is rnm-is sisxlSi

inficirebidan 1-2 kviris Semdeg aRmoCndeba (Thimme et al., 2001). HCV

hepatitis dros RviZlis ujredebis dazianebaSi did rols

TamaSobs Fas sistema (Hiramatsu et al., 1994). Fas aris ujredebis

zedapiruli proteini, romelic iwvevs apoptozs (Mita et al., 1994).

msoflioSi RviZlis cirozis da postransfuziuli hepatitis

ganviTarebis mizezad SemTxvevTa 90%_Si C hepatiti gvevlineba

posttransfuziuli da sayofacxovrebo gziT HCV-inficirebulTa

gamokvleviT dadginda, rom inficirebulTa 67%-s 6 Tvis Semdeg

alaninaminotransferazas momateba aReniSneba (Alter et al., 1995).

pirebs, romelTac Sratis transaminazebis aqtivobis maRali done

da anti- HCV zrda aReniSnebodaT, sisxlSi C hepatitis gamomwvevi

virusi (HCV-is rnm) aRmoaCndaT. am dros SeiZleba adgili hqondes

hepatocitebis aranormalur proliferacias, rac, Tavis mxriv,

hepatocelularuli karcinomis ganviTarebis risks zrdis.

amJamad ufro da ufro meti monacemebi metyvelebs imaze, rom

HCV-s qronizaciaSi did rols TamaSobs imunuri meqanizmebi (Wegstal

et al., 1995). arsebobs mtkicebuleba, rom qronikuli C hepatitis dros

adgili aqvs RviZlismier citotoqsiurobas da imunuri pasuxis

ganviTarebas, magram gaurkvevelia, Tu ra rols asrulebs igi

rogorc damcavi faqtori da rogorc qronikuli infeqciis

gamomwvevi faqtori (Nelson et al., 1997; Wong et al., 1998). am dros

matulobs IFNγ da iNOS-is genis eqspresia, romlebic monawileobas

iReben autokrinul da parakrinul procesebSi da xels uwyoben

qronikuli C hepatitis paTobiologias (Schweyer et al., 2000).

qronikuli HCV-s dros citoproliferacia ufro mkveTria, maSin

roca apoptozi imaze naklebia, vidre RviZlSi sxva dazianebebis

dros. disbalansi citopoliferaciasa da apoptozs Soris RviZlis

20

dazianebasa da RviZlis qsovilovani rkinis donesTan aris kavSirSi

(Farinati et al., 2001).

daavadebis simZimesa da viremias Soris arsebobs korelacia,

rac sisxlis SratSi alaninaminotransferazas aqtivobis momatebiT

dasturdeba. amas HCV-rnm-is titris mateba erTvis Tan. am dros

imunodepresiuli mkurnaloba, marTalia, aqveiTebs Sratismieri

transaminazebis aqtivobas, magram viremia matulobs (Kagawa et al.,

1993).

qronikuli C hepatitis dros Sratis transaminazebis

momateba RviZlis dazianebis xarisxs ar asaxavs. magram, Tu igi 10-

jer aRemateba normas, maSin es anTebiTi da nekrozuli

cvlilebebis momatebis mauwyebelia (Haber et al., 1995).

qronikuli C hepatitis dros SesaZlebelia, autoimunuri

hepatitic ganviTardes, rac zogierTi avtoris azriT,

interferoniT aris inicirebuli (Garcia-Buey et al., 1995).

autoimunuri hepatitisTvis damaxasiaTebelia SratSi

transaminazebis aqtivobis da autoantisxeulebis titris uecari

momateba. am dros imunodepresiuli Terapia efeqturia (Sherlock,

1999).

virusuli hepatitebis dros RviZlis dazianebaSi oqsidant-

antioqsidanturi balansis darRveva mniSvnelovan rols TamaSobs

(Satoh et al., 1996). am dros hepatocitebSi grovdeba didi raodenobiT

Tavisufali rkina, rac azianebs RviZlis membranul da

submembranul struqturebs. amasTanave aqtiurdeba lipidebis

zeJanguri Jangva, inaqtivirdeba mitoqondriuli

superoqsiddismutaza da viTardeba oqsigenuri stresi (Schwarz et al.,

1996). lipidebis zeJanguri Jangva Jangbadis Tavisufali

radikalebis warmoqmnis klasikur gzas warmoadgens. igi

fosfolipidebis membranis struqturis Secvlas da misi

mTlianobis darRvevas ganapirobebs, Tavisufal-radikaluri

21

saxeobebi membranis dazianebiTa da transmembranuli ionuri

balansis SecvliT ujredebis sikvdils ganapirobeben (Halliwell et al.,

1990). enzimuri da araenzimuri oqsidanturi sistemis mravali

komponenti (superoqsiddismutaza, katalaza, glutaTion

peroqsidaza, aRdgenili glutaTioni, fosfolipidebi)

dainhibirebuli arian toqsinebis mier, amave dros sisxlSi

matulobs ceruloplazminis done (Korda, 1996).

oqsigenuri stresi, Tavis mxriv, iNOS-is gaaqtiurebas da

azotis oqsidis Warb produqcias iwvevs (Turpaev et al., 1998). iNOS-is

produqcia izrdeba hepatocitebSi stresis warmoqmnis sapasuxod. es

pasuxi ki xorcieldeba mraval sxvadasxva faqtors Soris

kompleqsuri urTierTobiT, rogorebicaa kupferis ujredebis

gaaqtiureba, reaqtiuli Jangbadis formebis warmoqmna,

kolagenozebis warmoqmnis endotoqsiuri kontaminacia (Tirmenstein et

al., 2000). NO-s moqmedebis erT-erTi Sedegi ki mis mier

antioqsidanturi fermentebis aqtivobis gazrda (Dobash et al., 1997) da

am fermentTa makodirebuli genebis eqspresiaa (Nunoshiba et al., 1993).

garda amisa, Tavad NO antioqsidantur Tvisebebsac amJRavnebs.

dadgenilia, rom NO TNFα-iT inducirebuli kaspaza 3 da kaspaza 8-

is gaaqtiurebisagan da ujredis apoptozuri signalisagan

hepatocitTa dacvas uzrunvelyofs (Li et al., 1999; Rai et al., 1998). NO

xels uwyobs IL-10-is simsivnis sawinaaRmdego moqmedebas (Sur et al.,

2000).

unda aRiniSnos isic, rom NO-s citoproteqtorul

zemoqmedebasTan erTad, rasac igi apoptozis daTrgunviT aRwevs,

amave ujredebis sikvdilis gamowvevac SeuZlia (Melino et al., 1997). NO

Trgunavs NF-kB-s eqspresias endoTelur ujredebSi (Spiecker et al.,

1998). qronikuli virusuli hepatitebis mimdinareobaSi NO

mniSvnelovani paTologiuri procesebis Camoyalibebas ganapirobebs

(Majano et al., 1998). paTologiuri procesebi ki IFNα, TNFα, baqteriuli

22

polisaqaridiT da deqsametazoniT regulirdeba. amasTan, IFNα

iwvevs iNOS-is genis eqspresias, TGF-β1 ainhibirebs citokin-

inducirebuli NO-s gamomuSavebas iNOS-is post-transfuziuli

sinTezis blokirebiT (Koide et la., 1994).

RviZlis imunuri dazianebis meqanizmebis Sesaswavlad

eqsperimentuli modelebi gamoiyeneba. konkanavalin A (Con-A)-Ti

gamowveuli hepatiti, rogorc RviZlis imunuri dazianebis

eqsperimentuli modeli, farTod gamoiyeneba hepatitebis

paTogenezisa da mkurnalobis axali gzebis Sesaswavlad (Tiegs et al.,

1992; Tagawa et al., 1997; Nikoletti et al., 2000; Kato et al., 2001). Con-A aris

leqtini, romelic aaqtivebs limfocitebsa da monocitebs,

romlebic, Tavis mxriv, in vitro gamoimuSaveben sxvadasxva citokins.

igi RviZlis sinusis mimarT maRali afinurobiT xasiaTdeba (Tiegs et

al., 1992). Con-A-Ti gamowveuli RviZlis dazianeba T ujredebis

gaaqtiurebis Sedegad gamowveuli hepatocitebis apoptozisa da

nekrozis Sedegia. Con-A-s akumulacia RviZlis sinusebSi

mocirkulire limfocitebis gazrdil modenas da Semdgom,

blastoiduri formaciis gziT mis lokalur proliferacias iwvevs.

RviZlSi imunuri ujredebis gaaqtiureba iwvevs im citokinebis

momatebul eqspresias, romlebic RviZlis damazianebeli efeqtiT

xasiaTdebian (Gantner et al. 1995; Leist et al., 1995). limfocitebis

infiltracia RviZlis qsovilSi ukve Con-A–s Seyvanidan 8 saaTis

Semdeg iwyeba, xolo citokinTa umravlesobis maqsimaluri done

limfocitebis infiltraciamde miiRweva. es ki aCvenebs, rom

citokinebis donis adreuli momateba RviZlis ujredebis

dazianebaSi mniSvnelovan rols asrulebs (Mizuhara et al., 1994;

Trautwein et al., 1998). Con A-inducirebuli hepatitebis dros mkveTrad

izrdeba CD8 matriculi rnm, romelic citotoqsiuri T ujredebis

markeria, magram, CD8+ T-ujredul infiltracias minimaluri efeqti

gaaCnia hepatitebis ganviTarebaSi (Okamoto et al., 1998). am mxriv

23

kritikul mediatorebs interferoni (IFN) da simsivnis nekrozis

faqtori (TNF) warmoadgenen (Ksontini et al., 1998; Trautwein et al. 1998;

Tagawa et al., 1997). maTi doneebi momatebulia. TNF aris citokini,

romelic ZiriTadad gaaqtiurebuli makrofagebisgan da zogierTi

sxva tipis ujredis mier gamomuSavdeba. anti-TNF antisxeulebis

Seyvana Con-A–s daniSvnamde Tagvebs RviZlis dazianebisgan icavs,

rac ganpirobebuli TNF-s donis mniSvnelovani SemcirebiT.

TNF mniSvnelovani mediatoria sxvadaxva fiziologiuri da

paTologiuri mdgomareobis da apoptozis dros. igi aucilebelia

RviZlis regeneraciis dros hepatocitebis normaluri

proliferaciisTvis. TNF funqcionirebs, rogorc komitogeni da

ainducirebs birTvuli transkrifciis faqtor kB-s, romelsac

antiapoptozuri efeqti aqvs. meore mxriv, Con A-inducirebuli

hepatitis dros TNF hepatotoqsiurobis mediatoria, iwvevs ra

apoptozsa da nekrozs. igi agreTve mniSvnelovani mediatoria

virusuli hepatitebis dros (Ray et al., 1998). rac Seexeba IFN-s, igi

aadvilebs makrofagebis an NK ujredebis aqtivacias da

proliferacias (Guidotti et al., 2001), MHC II klasis eqspresias (Gantner

et al., 1995; Kustner et al., 1996). hepatocitebis dazianebas agreTve

ganapirobeben Fas/Fas ligandi da perforini (Iwai et al., 1999; Tagawa et

al., 1998).

histologiurad, Con-A inducirebuli hepatitis dros xdeba T

da polimorfobirTvuli ujredebis infiltracia portul areSi

da centraluri venis irgvliv. garda amisa, Con-A inducirebuli

hepatitisTvis damaxasiaTebelia zonaTaSorisi nekrozi. nekrozis

areSi aRmoCenilia CD4+ T limfocitebi (24 saaTis Semdeg), xolo 48

saaTis Semdeg rogorc CD4+, aseve CD8+ T limfocitebi (Satoh et al.,

1996).

Con-A inducirebuli hepatitis dros anTebis makontrolebeli

mniSvnelovani faqtoria agreTve IL-10 (Watanabe et al., 1996; Tiegs et al.,

24

1997; Louis et al., 2003). igi TNF-s da IFN-s gaaqtiurebis daTrgunvis

gziT RviZlis dazianebas amcirebs (Fiorentino et al., 1991). endotoqsinis

moqmedebidan maleve kupferis ujredebis mier IL-10-is genis

eqspresia xdeba, xolo mogvianebiT mas gamoimuSveben periferiuli

sisxlis ujredebi, magaliTad makrofagebi (Knolle et al., 1998). G zemoT

CamoTvlili citokinebis garda Con-A inducirebuli hepatitis dros

anTebis ganviTarebis mniSvnelovani citokinia IL-12 (Masayoshi et al.,

2000). garda amisa, am dros SeiniSneba citoqrom P-450-is genis

supresia (Okamoto, 1999).

unda aRiniSnos, rom Con-A damokidebuli hepatitis dros

citokinebis eqspresiaze pasuxismgebelni arian kupferis ujredebi

(Okamoto, 2000). am dros gamoiyofa IL-6, romelic damcvel efeqts

avlens (Mizuhara et al., 1994). garda amisa, sagrZnoblad matulobs IL-2,

IL-4-is doneebic (Masayoshi et al., 2000).

Con-A-s Seyvanis Semdeg sagrZnoblad matulobs ICAM-1 da

VCAM-1-is eqspresia (Watanabe et al. 1996), rac korelaciaSia

ujredebSi NF-kB Zlier da swraf gaaqtivebasTan. amrigad, SesaZloa

vifiqroT, rom NF-kB-s gaaqtiureba RviZlis dazianebaSi

monawileobs (Trautwein 1998).

Con-A-s Seyvana iwvevs ara marto RviZlis, aramed sxva

eqstrahepaturi organoebis dazianebas (elenTa, filtvebi,

sakvercxeebi). elenTaSi izrdeba IL-2-is done, xolo elenTaSi,

filtvebsa da sakvercxeebSi IFNγ–isa da TNFα-s Semcveloba

matulobs (Okamoto et al., 1998).

mravali Sroma mieZRvna Con-A inducirebuli hepatitebis

mkurnalobis Seswavlas. magaliTad, Con-A inducirebuli

hepatitebis dros aminoguanidini ainhibirebs

alaninaminotransferazebis aqtivobas, magram ar ainhibirebs

RviZlSi IL-2, IFNγ, TNFα da iNOS matriculi rnm-is eqspresias

(Okamoto et al., 2000).

25

LPS xels uSlis Con-A inducirebuli hepatitis ganviTarebas,

rac IL-6-is eqspresiis gaZlierebiT da NK1+ T ujredebis mier

sinTezirebuli IL-4-is da IFNγ-s inhibirebiT miiRweva. garda amisa,

LPS ainhibirebs IL-2Rα-s eqspresias (Nishikage et al., 1999).

Con-A inducirebuli hepatitebis dros RviZlis imunuri

dazianebis sawinaaRmdegod efeqturia dabalmolekuluri masis

heparinis gamoyeneba (Hershkoviz et al., 1999).

naCvenebi iqna, rom Con-A inducirebuli hepatitebis dros

beraprosti (prostaglandinebis ojaxis wevri) amcirebs RviZlis

dazianebas da amcirebs sikvdilobas, agreTve alt-s dones

plazmaSi da RviZlis histologiur suraTs aumjobesebs.

prostaglandinebi da misi analogebi mniSvnelovnad zrdis cikluri

adenozinmonofosfatis koncentracias prostaglandinis

receptorebis saSualebiT, rac Tavis mxriv aaqtiurebs

proteinkinaza A-s signalis transduqcias. am proteinkinazis

gaaqtiureba Trgunavs T ujredebis proliferaciasa da citokinebis

warmoebas NF-κB–is transkripciuli aqtiurobis daTrgunvis

saSualebiT, romelic anTebiTi citokinebis mTavar transkripciul

faqtors warmoadgens. marTalia, beraprosti zrdis splenocitebis

mier interferon-gamas produqcias in vitro pirobebSi, in vivo piriqiT

interferonis produqciis sarwmuno inhibireba xdeba.

vinaidan hepatitebis dros vlindeba oqsidaciuri stresi,

erT-erT Terapiul saSualebad ganixileba iseTi preparatebis

gamoyeneba, romlebic moqmedeben oqsidaciur procesebze.

Seswavlili iqna egzogenuri azotis oqsidis donoris

molsidominis gavlena Con-A inducirebuli hepatitis modelze.

aRmoCnda, rom citoproteqtoruli meqanizmi drozea damokidebuli.

egzogenuri azotis oqsidis donoris Seyvana RviZlis dazianebamde

(profilaqtikuri mizniT) an dazianebis Semdeg RviZls hepatitis

Sedegad ganviTarebuli cvlilebebisgan icavs. molsidominiT

26

mkurnaloba Con-A inducirebidan Tundac sami saaTis Semdeg

RviZls dazianebisgan icavs da rekomendebulia misi klinikaSi

gamoyeneba (Ding et al., 2004).

glicirizini xels uSlis Con-A inducirebuli hepatitis

ganviTarebas efeqturi citokinebis eqspresiis gareSe (Okamoto et al.,

1999).

27

1.2 apoptozis roli RviZlis normalur funqcionirebaSi

zrdasruli RviZlis parenqimaSi ujredebis gamravlebisa da

apoptozis tempebis Sesaxeb informacia mwiria. fiziologiurad

apoptozi Zalian Znelad fiqsirdeba, radgan ujredebis cvlis

masStabi minimaluria. RviZlSi yovel aTi aTas ujredze mxolod 1-

5 apoptozuri ujredis dadgena xerxdeba (Schulte-Hermann et al., 1995).

odnav gansxvavebulia es maCvenebeli dieturi faqtorebis

zemoqmedebis dros (Albright et al., 1997).

gare Tu Sida gzebiT warmoebuli apoptozi arsebiTia

RviZlSi ujreduli homeostazis SenarCunebaSi (Igney et al., 2002). gare

gza ujredis zedapiridan Fas-iT, TNF–iT an TGF--iT gaiSveba. Sida

gza iwyeba mitoqondriidan. masze zegavlenas axdens Bcl-2 ojaxis

wevrebi. orive gza urTierTgadaxlarTulia da gamorCeul

fiziologiur rols TamaSobs RviZlSi. apoptozuri gzebis

regulaciis moSla xels uwyobs iseTi daavadebebis ganviTarebas,

rogoricaa hepatocelularuli karcinoma, virusuli hepatiti,

autoimunuri hepatiti, iSemia. apoptozis defeqti xSirad

centraluri paTogenetikuri movlenaa. aqedan gamomdinare,

apoptozis molekuluri meqanizmebis Seswavla ara marto

daavadebebis meqanizmebis Sesaxeb sruli suraTis CamoyalibebisTvis

aris aucilebeli, aramed xels uwyobs RviZlis daavadebis

potenciuri Terapiuli meTodebis SemuSavebis process.

1.2.1 gare apoptozuri gzebi

Fas prototipiuri sikvdilis receptoria. is pirvelad 1989

wels aRweres (Trauth et al., 1989; Yonehara et al., 1989) da warmoadgens 42-52

kilodaltoni molekuluri wonis mqone glikozilirebul I tipis

transmembranul proteins (ujredgare aminoterminaluri domeni).

Fas RviZlis yvela ZiriTadi saxis ujredzea eqspresirebuli

(hepatocitebi, stelaturi ujredebi, kupferis ujredebi da RviZlis

28

endoTeluri ujredebi). is Sesabamisi specifiuri ligandiT

aqtivirdeba. Fas-is liganda II tipis transmembranuli proteinia

(ujredSida aminoterminaluri domeni). Fas-sgan gansxvavebiT, misi

liganda RviZlSi fiziologiurad ar aris eqspresirebuli, magram

aqtiurdeba RviZlis dazianebis SemTxvevaSi (Eichhorst et al., 2000). Fas-

isa da misi ligandis koeqspresia ki ujredis TviTmkvlelobiT

mTavrdeba. garda amisa, ligandas apoptozis gamowveva

eqspresirebuli Fas-is mqone mezobel ujredebSic SeuZlia

(parakrinuli sikvdili).

Fas-is mTavari funqciaa apoptozis procesis gaSveba (Krammer et

al., 1994). marTalia, is sxvadasxva qsovilsa da ujredul xazSi

gvxvdeba (Moller et al., 1994; Weller et al., 1994), misi upiratesi

fiziologiuri roli SesaZloa imunuri sistemis regulaciaSi

mdgomareobs. amis magaliTia MRL/lpr Tagvis fenotipi, romelic Zlier

gaaqtiurebuli Fas-is eqspresias aCvenebs (Mariani et al., 1994). Lpr Tagvs

uviTardeba limfadenopaTia da splenomegalia, garda amisa,

gamoumuSavdeba auto-antisxeulebis didi raodenobebi. Fas-is

molekula Tavdapirvelad miCneul iqna rogorc ujredis zedapiruli

receptori, romelsac transformirebuloi ujredebis apoptozuri

sikvdilis mediatoroba da SiSvel TagvebSi mzardi eqsperimentuli

simsivneebis regresia SeeZlo (Trauth et al., 1989; Yonehara et al., 1989).

seriozuli sistemuri efeqtebis miuxedavad, Fas-iT warmoebuli

apoptozis gamoyenebam imedebi ar gaamarTla kibos TerapiaSi. meore

mxriv, dReisaTvis ukve mravali dokumentirebuli monacemi arsebobs,

romlebic ujredis sikvdilis sxvadasxva fiziologiur da

paTologiur formebSi Fas-is arsebiT rolze miuTiTebs (Nagata et al.,

1995; Schulze-Osthoff et al., 1994). hepatocitebi mudmivad axdenen Fas-is

eqspresias TavianT zedapirze. naCvenebia, rom TagvebSi am receptoris

stimulireba ramdenime saaTSi RviZlis muSaobis fulminantur

ukmarisobas iwvevs (Ogasawara et al., 1993; Rensing-Ehl et al., 1995). vilsonis

29

mwvave daavadebis dros RviZlis ukmarisoba da RviZlis ujredebis

dazianeba moicavs Fas-iT ganviTarebul apoptozs (Strand et al., 1998).

Fas/Fas-ligandis sistemis dadebiTi regulacia demonstrirebulia

alkoholur cirozSi, qronikul virusul hepatiti B-sa da C-Si da

gansxvavebuli etiologiis mqone RviZlis fulminantur ukmarisobaSi

(Strand et al., 1998).

Fas-sa da TNF-is receptor 1-s gaaCniaT erTnairi mokle

ujredSida domeni - e.w. “sikvdilis domeni” (Tartaglia et al., 1993).

membranebTan dakavSirebuli ligandis ararsebobis dros yalibdeba

Fas-is inaqtiuri kompleqsi (Scaffidi et al., 1998). Fas Tavis ligandasTan

dakavSirebis dros apoptozur signals ujredSi agzavnis e. w.

“sikvdilis domenis” saSualebiT. Tumca, signalis efeqturi

gadacemisTvis saWiroa agreTve e.w. “sikvdilis gamomwvevi

signalirebis kompleqsis” Camoyalibeba. es aris Fas receptorTan

dakavSirebuli ujredSida proteinuli kompleqsi, romelic

ujredis SigniT erT-erTi Tavisi komponentis “Fas-asociirebuli

sikvdilis domenis” saSualebiT ikavSirebs prokaspaza 8-s. Tavis

mxriv, prokaspaza 8 ganicdis avtolizs. ori prokaspaza 8-is patara

da didi domenebidan warmoiqmneba erTi aqtiuri kaspaza 8 (ori

didi da ori patara domeniT), romelic iniciatori kaspazaa, radgan

is kaskadurad aaqtiurebs yvela danarCen efeqtor kaspazas da

saboloo jamSi ujredis apoptozur sikvdils iwvevs (Cohen et al.,

1997). efeqtori kaspazebis aqtivirebis garda, kaspaza 8-s SeuZlia

Bcl-2 ojaxis wevris Bid-is gaxleCva da Semoklebuli Bid-is (tBid)

warmoqmna. es ukanaskneli ki aaqtiurebs mitoqondrias, riTac

kavSiri myardeba sikvdilis receptoriT signalirebasa da Sida

apoptozur gzas Soris.

Fas ligandasTan urTierTqmedeba am kompleqsis reorganizebas

axdens, rasac moyveba “sikvdilis gamomwvevi signalirebis

kompleqsis” formireba. igi Sedgeba “Fas-asociirebuli sikvdilis

30

domenis adaptori proteinebisgan”, kaspaza 8 da 10-isgan, romlebsac

SeuZliaT apoptozis iniciireba. Fas-is ligandis mier “sikvdilis

gamomwvevi signalirebis kompleqsis” farglebSi Fas-is, “Fas-

asociirebuli sikvdilis domenis adaptori proteinebis”, kaspaza 8-

isa da kaspaza 10-is dajgufeba iwvevs mocemuli kaspazebis

autoproteolizur damuSavebasa da aqtiuri proteazebis

gamoTavisuflebas. I tipis ujredebSi damuSavebuli kaspaza-8

sakmarisia kaspazebis ojaxis sxva wevrebis gasaaqtiureblad,

romelTa mier Sesabamis substratebze zemoqmedeba apoptozis

egzekuciur fazas uxsnis gzas (Krammer et al., 2000). II tipis ujredebSi

Fas-is mier efeqtori kaspazebis saTanado gaaqtiureba damokidebulia

amplifikaciur maryuJze, rac Tavis mxriv damokidebulia kaspaza-8-is

mier Bcl-2 ojaxis proapoptozuri wevris Bid-is gaxleCvaze, rogorc es

aRwerilia TNF-is receptori 1-is sistemis SemTxvevaSi. Semdeg etapze

mitoqondriebi gamoaTavisufleben proapoptozur faqtorebs,

magaliTad, citoqrom c-s, Diablo-sa da “apoptozis gamomwvev fators”,

romlebic ganapirobeben kaspaza-9-is gamaaqtiurebeli proteinuli

kompleqsis Semcveli apoptosomis Camoyalibebas. apoptosoma Sedgeba

citoqromi c-is, APAF-1-is, dATP-isa da pro-kaspaza-9-isgan.

gaaqtiurebuli kaspaza-9 aaqtiurebs aRmasrulebel kaspaza-3-s,

kaspaza-3 ki - kaspaza-8-s, riTac ikvreba kidec dadebiTi pasuxis

maryuJi (Krammer et al., 2000). am ori ZiriTadi gzis garda,

SemoTavazebulia kidev aranakleb ori gza, romlebic Fas-is

signalirebas awarmoeben.

kidev erTi adaptori molekula – “receptorTan

urTierTmoqmedi proteini (anu RIP)” - ayalibebs kompleqss “RIP-Tan

dakavSirebeli interleikin-1β-gardamqmneli fermentis msgavsi

proteaza 1-is homologiur sikvdilis domenTan” da kaspaza-2-Tan da

Fas-is signals gadascems kaspaza-8-is sawyisi gaaqtiurebisgan

damoukideblad (Duan et al., 1997). FADD-isa da RIP-is garda, aRwerilia

31

Fas-is makavSirebeli kidev erTi proteini - “Fas-is sikvdilis

domenTan asociirebuli proteini” - romelic ayalibebs kompleqss

“apoptozis signalis maregulirebel kinaza 1-Tan”, rasac moyveba “c-

Jun aminoterminaluri kinazisa” da mitogeniT aqtivirebuli protein

kinaza p38-is gaaqtiureba. p38-s Tavis mxriv SeuZlia transkrifciis

faqtoris - E2F1-is - gaaqtiureba, romelic miCneulia S-fazasa da

kaspazebTan dakavSirebuli proteinebis makodirebeli genebis

erTdrouli transkrifciis gamomwvevad (Wang et al., 1999; Kornmann et al.,

2000). Tumca, axlaxans vilungerisa da misi kolegebis kvlevis

Sedegebma kiTxvis niSnis qveS daayena apoptozur signalirebaSi RIP-

isa da “Fas-is sikvdilis domenTan asociirebuli proteinis”

monawileoba (Villunger et al., 2000). Fas-is mier warmoebuli apoptozis

ramdenime etapi SeiZleba iyos regulatoruli meqanizmebis samizneebi,

rac saSualebas aZlevs ujredebs Fas-is mier stimulirebas moqnilad

upasuxon. “sikvdilis gamomwvevi sasignalo kompleqsis” mier kaspaza-

8-is gaaqtiurebis unari ZiriTadad regulirdeba “FADD-is msgavsi

inteerleikin-1β-gardamqmneli fermentis msgavsi mainhibirebeli

proteinis mier” (Krueger et al., 2001). es proteini ramdenime izoformiT

arsebobs. es izoformebi struqturulad kaspaza-8-is msgavsia, Tumca

ar gaaCniaT fermentuli aqtiuroba (Krueger et al., 2001; Irmler et al., 1997).

igi SeiZleba CarTuli iyos “sikvdilis gamomwvevi sasignalo

kompleqsis” sikvdilis receptorebSi, riTac is aCerebs kompleqsis

mier kaspaza-8-is procesingsa da misi gaaqtiurebuli formis

gamoTavisuflebas. garda amisa, II tipis ujredebSi Fas-is mier

warmoebuli apoptozi kontrolirdeba ujredis sikvdilis

mitoqondriuli gzis mravali regulatoriT, kerZod, Bcl-2 ojaxis

wevrebis mier (Krammer et al., 2000). Fas-is mier warmoebuli ujredis

sikvdili xdeba ara marto apoptoziT, aramed - ujredul konteqstis

mixedviT – nekroziTac (Leist et al., 1997), romelic saWiroebs adaptor

protein FADD-sa da Fas-Tan urTierTmoqmed serin/treonin kinaz

32

receptorTan urTierTmoqmed proteins, magram ara - kaspaza-8-s (Holler

et al., 2000). imavdroulad, isic unda aRiniSnos, rom nekrozis

egzekuciur procesTan Fas-is damakavSirebeli meqanizmebi jer ar

aris garkveuli.

1.2.1.1 simsivnis nekrozis faqtoris gza

TNF anTebisa da imunurobis mTavari mediatoria. garda amisa,

is iwvevs RviZlis mwvave fazis reaqcias, RviZlis regeneraciasa da

zrdas, imunuri ujredebis stimulirebas, adheziuri molekulebis

dadebiT regulacias, Sokismagvar mdgomareobas, lipolizs, kaxeqsias,

qsovilis destruqciasa da ujredis apoptozur Tu nekrozul

sikvdils ujredis sxvadasxva tipSi (Aggarwal et al., 1992; Tracey et al.,

1993). ufro metic, TNF CarTulia adamianis sxvadasxva daavadebis, maT

Soris, sefsisis, diabetis, kibos, osteoporozis, gafantuli

sklerozis, revmatoiduli arTritis paTogenezSi.

TNF ujreduli efeqtebi gadaicema ori specifiuri zedapiruli

receptoriT: TNF-R1 (20 kilodaltoni) da TNF-R2 (75 kilodaltoni)

(Leist et al., 1995; Leong et al., 2000). TNF-isa da TNF-R1-is urTierTmoqmedeba

aaqtiurebs signalis transduqciis ramdenime gzas. am gzebis saerTo

niSania TNF-is mier ujredis membranaze signalirebis

mravalproteiniani kompleqsis, e.w. “sikvdilis gamomwvevi

signalirebis kompleqsis” Camoyalibebis gamowveva. erTi gza

sruldeba kaspazebis gaaqtiurebiT (Sidoti-de Fraisse et al., 1998), meore -

neitraluri sfingomielinazis mier ceramidis warmoqmniT, mesame

xasiaTdeba “c-Jun aminoterminaluri kinazis” aqtivaciiT da

transkrifciis faqtori c-Jun-is Tanamdevi gaaqtiurebiT, meoTxe gzas

ki apoptozis Semaferxebeli transkrifciis faqtori NF-B-is

aqtivaciamde mivyavarT (Liu et al., 1996). transkrifciis aRniSnuli

faqtorebi pasuxismgebeli arian sxvadasxva biologiuri procesis,

maT Soris, ujredis zrdisa da sikvdilis, ganviTarebis, onkogenezis,

33

imunuri, anTebiTi da stresuli pasuxebisaTvis mniSvnelovani genebis

eqspresiis gamowvevaze. am gzebis balansi ujredis beds gadawyvets.

sqema. 2. TNF signalis transduqciis gza. TNF-is dakavSireba mis monaTesave

receptorebTan iwvevs “sikvdilis domenis inhibitoris” Camocilebasa da

sikvdilis gamomwvevi signalirebis kompleqsis Camoyalibebas, romelic

Sedgeba “TNF-is R1 receptorTan asociirebuli sikvdilis domenisgan”,

“TNF-is receptorTan asociirebuli faqtori-2-gan” da RIP-isgan. Semdgomi

movlenebi iniciirdeba IκB kinazis IKKβ suberTerulis kompleqsiTa da

kaspaza-8-iT

TNF-is signalis gadacemis procesis sawyisi moicavs TNF-R1-is

ujredgare domenTan TNF-is trimeris dakavSirebasa da TNF-R1-is

ujedSida sikvdilis domenisagan mainhibirebeli proteinis, e.w.

sikvdilis domenebis sailenseris Camocilebas (Tschopp et al., 1999; Jiang

et al., 1999). warmoqmnili agregirebuli receptori amoicnoba

adaptoruli proteinis, e.w. TNF-is receptorTan asociirebuli

sikvdilis domenis mier, romelic moizidavs damatebiT adaptorul

34

proteinebs, rogoricaa “receptorTan urTierTmoqmedi proteini”,

“TNF-is receptorTan asociirebuli faqtori 2” da “Fas asociirebuli

sikvdilis domeni” (Hsu et al., 1996; Stanger et al., 1995; Chinnaiyan et al., 1995).

es proteinuli kompleqsi pasuxs agebs signalirebis movlenebis

dawyebaze (Rath et al., 1999)

1.2.2 Sida apoptozuri gza

Sida apoptozuri gza ujredis Signidan stimulirdeba. misi

gamomwvevi SeiZleba iyos ultraiisferi gamosxiveba, reaqtiuli

Jangbaduri saxeobebi (ROS), zrdis faqtoris mocileba an

qimioTerapiuli wamlebi). am dros Bcl-2 ojaxis wevrebis - Bak-isa

da Bax-is - saSualebiT mitoqondriidan iwyeba kaspazaze

damokidebuli da damoukidebeli apoptozuri gzebi.

1.2.2.1 apoptozis mitoqondriuli regulireba

mitoqondriuli kaspazis aqtivacia mWidro kavSirSia

mitoqondriis gare membranis gamtarobis gazrdasTan (Green et al.,

2004). am procesis regulaciaSi CarTulia Bcl-2 ojaxis proteinebi,

mitoqondriuli lipidebi, bioenergetikuli metabolitebis

moZraobis maregulirebeli proteinebi da mitoqondriis membranaSi

arsebuli mitoqondriuli gamtarobis makotrolebeli foris

komponentebi, maT Soris, Zabva-damokidebuli anionuri arxi, adenin

nukleotid transportiori da am transportiorTan

urTierTmoqmedi proteini ciklofilini D. mitoqondriis gare

membranis gamtarobis gazrda iwvevs intermembranuli sivrcis

proteinebis, kerZod, citoqrom c-is, prokaspaza 9-is, “apoptozis

gamomwvevi faqtoris”, “kaspazis mitoqondriidan warmoSobili meore

aqtivatorisa” da Htra2/Omi-s gamoTavisuflebas. am procesis zusti

meqanizmi jer-jerobiT ucnobia da intensiuri kvlevis procesSia.

Bcl-2 ojaxis wevrebisa da baqteriuli toqsinebis forebis Semqneli

35

spiralebis struqturuli msgavsebis safuZvelze SemoTavazebulia

hipoTeza (Mucmore et al., 1996), rom Bcl-2 ojaxis wevrebi - Bak da Bax -

mitoqondriis gare membranaSi forebs qmnian, rac ganapirobebs

intermembranuli sivrcis proteinebis gamoTavisuflebas. sxva

modelis mixedviT, Bcl-2 ojaxis wevrebi SesaZloa urTierTqmedebs

mitoqondriis specifiur proteinebTan da xels uwyobs gamtarobis

cvlilebas makontrolebeli forebis gaRebiT. mesame SesaZlo

scenaris mixedviT, Bcl-2 ojaxis wevrebi SesaZloa iwvevs

mitoqondriis membranis formisa da lipidis forebis Secvlas.

yvela hipoTeza gadaikveTeba intermembranuli sivrcis proteinebis

gamoTavisuflebaze. gamoTavisuflebuli citoqromi c prokaspaza 9-

sTan, citoplazmur protein Apaf-1-Tan da ATP-sTan erTad qmnis

mravalproteininan kompleqss, apoptosomas, rac, Tavis mxriv,

aaqtiurebs prokaspaza 9-s da Sesabamisad xels uwyobs ujredis

kaspaza-damokidebuli sikvdilis dawyebas.

gamoTavisuflebuli “apoptozis gamomwvevi faqtori”

apoptozs kaspazis gareSe iwvevs (Susin et al., 1996). am gziT gamowveuli

apoptozis damaxasiaTebeli niSania dnm-is fragmentacia did

fragmentebad (daax. 50 kilodaltoni). “kaspazis mitoqondriidan

warmoSobili meore aqtivatori” da Htra2/Omi kaspazis aqtivaciis

fizilogiurad Semaferxebeli “apoptozis mainhibirebeli

proteinebis” inhibirebas axdenen. amrigad, “kaspazis mitoqondriidan

warmoSobili meore aqtivatori” da Htra2/Omi arapirdapiri gziT

uwyoben xels kaspazis gaaqtiurebasa da kaspaza-damokidebuli

apoptozis ganviTarebas.

36

1.3 kaspazebis roli apoptozSi

apoptozi koordinirdeba cisteinis proteazebis ojaxiT - e. w.

kaspazebiT - romlebic proteinebis mTeli arsenalis SezRuduli

proteolizis gziT ujredis demontaJs awarmoeben. ZuZumwovarTa

kaspazebis ojaxi 14 wevrisgan Sedgeba, romelTa nawili sakuTriv

apoptozSi monawileobs, danarCeni wevrebi ki savaraudod anTebis

citokinebis procesingSi monawileobs. kaspazebis apikaluri

aqtivaciis movlenebi Cveulebriv iniciirdeba adaptorebi

molekulebis mier, romlebic ganapirobeben kaspazebis agregacias da

xels uwyoben maT autoaqtivacias. maSin, roca kaspazebis distaluri

aqtivaciis movlenebi kontrolirdeba manamde kaskadSi ukve

gaaqtiurebuli kaspazebis mier, mravali ujreduli stresi apoptozs

iwvevs mitoqondriis dazianebiTac, romlis drosac

gamoTavisufldeba iseTi citokinebi, rogoricaa citoqromi c da

“kaspazis mitoqondriidan warmoSobili meore aqtivatori”. es

citokinebi ki, Tavis mxriv, aaqtiureben kaspazebs da ujredis

sikvdils iwveven.

kaspazebi ujredis sikvdilis meqanizmis Semadgenel

komponentebad pirvelad naCvenebi iqna Caenorhabditis elegans nematod

Wiaze warmoebul kvlevebSi. ujredis programirebuli sikvdilis

uunaro mutanti Wiebis genetikuri analizis saSualebiT dadginda,

rom ced-3 genis produqti saWiro yofila ganviTarebasTan

dakavSirebuli ujredis programirebuli yvela sikvdilis

ganxorcielebisTvis. Yuan-isa da misi kolegebis 1993 wlis

mniSvnelovani statiis mixedviT (Yuan et al., 1993), CED-3 geni mWidrod

ukavSirdeba adamianis proteaza interleikin-1 gardamqmnel

ferments. marTalia, aRmoCnda, rom es fermenti ar asrulebs

centralur rols apoptozSi, massa da CED-3-s Soris homologiis

Sesaxeb aRniSnul sawyis informacias swrafad mohyva am fermentis

msgavsi proteazebis aRmoCena, romlebsac SemdgomSi sxva

37

saxelwodeba - “kaspazebi” (cisteinis aspartat-specifiuri

proteazebi) – daerqva (Thornberry et al., 1998; Cohen et al., 1997; Earnshaw et

al., 1998). kaspazebi ZiriTad specifiurobas aspartamis mJavis

narCenebis mimarT amJRavneben, rac metad uCveulo substratis

mimarT mizidulobis magaliTia (Earnshaw et al., 1998; Green et al., 1998;

Martin et al., 1995). mocemuli fermentebi SedarebiT araaqtiuri

winamorbedebis (pro-kaspazebis) saxiT arseboben. maTi aqtiur

formaSi gardaqmna Sida aspartamis mJavis narCenebis proteolizuri

gaxleCvis gziT mimdinareobs. es miuTiTebs imaze, rom kaspazebi

autoproteoliziT an kaskadis zeviT gaaqtiurebuli sxva

kaspazebiT aqtiurdebian. kaspazebi zogadad SeiZleba daiyos or

funqciur jgufad: isini, romlebic apoptozis dros aqtiurdebian

(kaspaza-2, -3, -6, -7, -8, -9 da -10), da isini, romlebic imunuri pasuxis

dros anTebis citokinebis procesingSi arian CarTuli (kaspaza-1, -4,

-5 da -11). kaspazebi, romlebic apoptozSi monawileoben, Semdgom or

qvejgufad iyofa apoptozTan asociirebul kaspazur kaskadSi

poziciis mixedviT. zeda anu iniciatori kaspazebi (kaspaza-2, -8, -9 da

-10) zogadad apoptozis dros kaspazebis aqtivaciis kaskadis

iniciatorebad arian miCneuli. maT Cveulebriv grZeli prodomenebi

gaaCniaT, romlebic Seicaven protein-proteinuli kavSirebis mqone

monakveTebs, e.w. “kaspazebis mozidvis domenebs” an “sikvdilis

efeqtor domenebs”. es monakveTebi gvxvdeba agreTve kaspazebis

adaptor molekulebSi, rogoricaa FADD, Apaf-1, romlebic

apoptozis dawyebisas kaspazebis agregaciisa da aqtivaciis

amocanebs asruleben. meore jgufs, qveda anu efeqtor kaspazebs

(kaspaza-3, -6 da -7), romlebic apoptozis dros ujredis dangrevaze

arian uSualod pasuxismgebeli, prodemenebi imdenad ar gaaCniaT.

mraval sistemaSi kaspaza-3 mTavari aRmasrulebeli kaspazaa,

kaspaza-6 da kaspaza-7 ki SedarebiT mcire rols TamaSoben

apoptozis aRsrulebis fazaSi (Slee et al., 1999). aqtiuri kaspazebi

38

ujredis daSlas ramdenime gziT awarmoeben: garkveuli

proteinebis, magaliTad, Bid-is saSualebiT citoqrom c-s

gamoTavisuflebiT, sxva damangreveli fermentebis, maT Soris, dn-

azebis gaaqtiurebiT, ujredSi struqturul da regulatorul

proteinebze pirdapiri degradaciuli zemoqmedebiT. dReisaTvis

dadgenilia kaspazis SuamavlobiT substratis gaxleCvis ramdenime

mniSvnelovani movlena, romlebsac Tavisi wvlili SeaqvT

apoptozis fenotipSi. magaliTad, “kaspaziT aqtivirebadi

deoqsiribonukleaza” apoptozis dros internukleosomaluri dnm-

is fragmentaciaze pasuxismgeblobisaTvis yvelaze damajerebeli

kandidatia (Enari et al., 1997). igi Cveulebriv inhibirdeba Tavis

specifiur inhibitorTan asociaciiT. Tumca, es inhibitori kaspaza-3-

is substratia da am ukanaskneliT misi proteolizuri gaxleCva

“kaspaziT aqtivirebadi deoqsiribonukleazas” gamoTavisuflebas

xdis SesaZlebels. gamoTavisuflebuli deoqsiribonukleaza Sedis

birTvSi da qromosuli dnm-is degradacias iwyebs (Enari et al., 1997).

SemoTavazebulia, rom kaspaza-3-is substratia kidev erTi proteini

- “Rho-sTan asociirebuli kinaza 1” - apoptozuri ujredis

SekumSvasa da membranis blebings warmarTavs (Coleman et al., 2001;

Sebbagh et al., 2001). rogorc Cans, kaspazebi sakuTari aqtiurobis

gaZlierebasac axdenen inhibitori molekulebis, magaliTad,

“apoptozis inhibitori proteinis” gaxleCvisa da inaqtivaciis gziT

(Martin et al., 1995). marTalia, dReisaTvis kaspazebis mravali sxva

substratia gamovlenili, apoptozis konteqstSi maTi gaxleCvis

mniSvneloba jer-jerobiT gaurkveveli rCeba. axlaxans naCvenebi

iqna ujredis sikvdilis meqanizmis kidev erTi axali komponenti,

romlis saxelwodebaa “kaspazis mitoqondriidan warmoSobili

meore aqtivatori” (Du et al., 2000). es aqtivatori mitoqondriaSia

lokalizebuli. citozolSi misi gamoTavisufleba apoptozis

gamowvevis Semdeg xdeba. am dros is ukavSirdeba “apoptozis

39

inhibitor proteinebs”, riTac ixsneba kaspazebis aqtiurobis

inhibireba (Du et al., 2000).

rac Seexeba FADD-s, is receptors cisteinur proteazebs –

kaspaza 8-sa da kaspaza-10-s - ukavSirebs da ganapirobebs am

ukanasknelTa TviTgaxleCvasa da gaaqtiurebas, rac, Tavis mxriv,

ganapirobebs proteazuli kaskadis gaSvebasa da, saboloo jamSi,

apoptozs (Rath et al., 1999). erTi mxriv, kaspaza-8 uSualod aaqtiurebs

efeqtor proteazebs - kaspaza-3, -6 da -7-s - romlebic, Tavis mxriv,

ujredis mraval proteins xleCen da ujredis sikvdils iwveven.

meore mxriv, efeqtori kaspazebis aqtivacia Zlierdeba

mitoqondriuli gziT. kaspaza-8-is mcire aqtiurobac ki sakmarisia,

rom Bid gaixliCos. warmoqmnili e.w. “Semoklebuli Bid“ (tBid)

gadaadgildeba mitoqondriebisken da jer-jerobiT arasrulad

garkveuli meqanizmis wyalobiT mitoqondriuli membranis

potencials cvlis, riTac xeli ewyoba “mitoqondriuli gamtarobis

gadasvlis forebis” Camoyalibebasa da mitoqondriuli funqciis

moSlas (mimoxilulva ix. Hengartner, 2000). amis Semdeg mitoqodriebidan

citozolSi gadadis citoqromi c, ukavSirdeba Apaf-1-sa da pro-

kaspaza-9-s, ayalibebs proteinul kompleqss, e.w. apoptosomas, romlis

Semadgeneli nawilia dATP. rac Seexeba kaspaza-9-s, is damuSavebas

ganicdis da aaqtiurebs qveda kaspaza-3-sa da kaspaza-7-s (Hengartner,

2000; Li et al., 1999). kaspazebis gaaqtiurebis aRniSnuli mitoqondriuli

amplifikaciuri maryuJi regulirdeba Bcl-2 ojaxis anti-apoptozuri

proteinebiT.

dabolos, TNF signalirebis qselis saintereso

damaxasiaTebeli niSania apoptozur, NF-B-isa da “c-Jun

aminoterminaluri kinazis” signalirebis gzebs Soris eqstensiuri

jvaredini urTierTobebi. NF-B-is moqmedebis ararsebobis

SemTxvevaSi, TNF-is mier gamowveuli apoptozis mimarT ujreduli

mgrZnobeloba izrdeba. NF-B-is gamiznuli aqtivacia apoptozisgan

40

dacvas uzrunvelhyofs, magaliTad, “apoptozis mainhibirebeli

ujreduli proteinebis” sinTezirebiT, romlebsac kaspazebis

aqtiurobis pirdapiri inhibireba SeuZliaT (Deveraux et al., 1999). es

mainhibirebeli proteinebi Seicaven sam domens, –romelTagan pirveli

ori efeqtori kaspaza-3, -6 da -7-is moqmedebis blokirebas axdenen,

mesame ki SerCeviTad ainhibirebs kaspaza-9-is moqmedebas (Deveraux et al.,

1999). saerTo jamSi, arsebobs ramdenime gamSvebi punqti, romlebic

sikvdilis receptorebis mediatorobiT warmoebul ujredul

sikvdils akontroleben.

1.4 apoptozis modulacia Terapiuli saSualebebiT

ujredis apoptozuri sikvdilis mkacri regulacia da

gadarCenis gzebi Terapiuli intervenciebis strategiebis

SemuSavebis SesaZleblobas iZleva. RviZlis sxvadasxva daavadebis

dros (virusuli, autoimunuri hepatiti, iSemiur-reperfuziul

dazianeba, toqsikuri dazianeba da sxva.) apoptozis Warbi

gaaqtiureba aRiniSneba, maSin roca Semcirebuli apoptozi gvxvdeba

RviZlis regeneraciuli zrdis an RviZlis simsivnebis SemTxvevaSi.

dReisaTvis ukve mravali Sroma mieZRvna RviZlis sxvadasxva

dazianebis dros apoptozze moqmedi axali Terapiuli saSualebebis

gamoyenebas. miuxedavad amisa, warmatebuli modulatorebis povna

Zalian Znelia da am mimarTulebiT kvlevebi intensiurad

grZeldeba.

garda amisa, mkurnalobis samizneebad mniSvnelovania

agreTve dro, Tu rodis unda moxdes apoptozis modulacia, radgan

RviZlis daavadebebi SeiZleba ganxilul iqnes rogorc

mravalsafexuriani procesebi. sxvadasxva safexuria: ujredis

sikvdilTan asociirebuli RviZlis qronikuli dazianeba, cirozi

da regeneracia, apoptozis inhibiciasTan dakavSirebuli simsivnis

formireba da RviZlis ukmarisoba. amrigad, erTidaimave daavadebis

41

sxvadasxva etapze apoptozis inhibirebas an gaaqtiurebas

gasxvavebuli Sedegebi mohyveba.

1.4.1 hepatocitebze mimarTuli Terapia

mwvave dazianebis dros hepatociis apoptozis inhibireba

SeiZleba sasargeblo iyos. antiapoptozuri intervenciebis

samizneebs Sorisaa kaspazebi (kaspazebis endogenuri an egzogenuri

inhibitorebis gamoyeneba) da mitoqondriis erTianobis SenarCuneba

(Bcl-2 ojaxis antiapoptozuri wevrebis gamoyeneba). miCneulia, rom

xSirad anTebissawinaaRmdego agentebi RviZlis mwvave dazianebis

dros am dazianebas amcireben. magram gaurkvevelia, Tu ramdenad

Seesabameba mocemuli strategia yvela paTologiur mdgomareobas.

magaliTad, baqteriuli infeqciiT gamowveul RviZlis mwvave

daavadebaSi anti-TNF Terapia RviZlis dazianebis Tavidan acilebas

uzrunvelyofs, magram samagierod mcirdeba baqteriis ganadgureba

da gadarCena (Moore et al., 2003). zogierTma anTebissawinaaRmdego

strategiam SeiZleba Seasustos citokinebis warmoeba da NF-kB–is

aqtivacia da amiT apoptozis mimarT hepatocitebis mgrZnobelobis

Semcireba moaxdinos (Schoemaker et al., 2004). RviZlis qolestatikuri

dazianebis mqone pacientebs xSirad mkurnaloben ursodeoqsiqolis

mJaviT, romelic erT-erTi naRvlis mJavaa da adamianis naRvlis

mJavebis 3%-s Seadgens. es nivTiereba Tavdapirvelad Savi daTvis

RviZlidan iqna miRebuli da uxsovari droidan gamoiyeneba CineTis

tradiciul medicinaSi RviZlis daavadebebis samkurnalod

(Paumgartner et al., 2002). axlaxans naCvenebi iqna, rom ursodeoqsiqolis

mJavis taurinis warmoebuli naRvlis mJavis mier gamowveuli

apoptozisgan icavs organizms, rasac axorcielebs mitoqondriul

membranaze pirdapiri zemoqmedebiTa da “mitogen-gaaqtivebuli

protein kinazebis” gaaqtiurebiT (Schoemaker et al, 2004).

42

1.4.2 kaspazebi da maTi inhibitorebi

Bcl-2 ojaxis garda, kaspazebis inhibitorebi, rogoricaa

“apoptozis mainhibirebeli proteinebis” ojaxis wevrebi, ujreds

apoptozuri sikvdilisagan kaspazebis kaskadis darRvevis gziT

icavs. in vitro pirobebSi “apoptosis mainhibirebeli ujreduli

proteinis” adamianis homologis Warbi eqspresia hepatocitebs

apoptozisgan icavs (Schoemaker, Ros et al., 2002). aRniSnuli ojaxis

wevrebi SerCeviT ainhibireben kaspaza 3-sa da kaspaza 9-s. maTi

aqtiurobis dablokva SeiZleba “kaspazis mitoqondriidan

warmoSobili meore aqtivatoriTa” da Omi/HtrA2-iT. amrigad,

ujredis beds “apoptozis mainhibirebeli proteinebis” ojaxis

wevrebTan kaspaza 3-is, kaspaza 8-is, kaspaza 9-is, “kaspazis

mitoqondriidan warmoSobili meore aqtivatorisa” da Omi/HtrA2-is

fardobiTi citozoluri koncentraciebis balansi wyvets.

imisda mixedviT, Tu ra tipis dazianebasTan gvaqvs saqme,

garkveuli kaspazebis inhibireba saTanado strategiad unda iqnes

miCneuli. baitma da misma kolegebma (Bajt et al., 2001) aCvenes, rom

marTalia peptidebis inhibitoris mier kaspaza 3-is inhibireba

aferxebda LPS/GalN–iT (d-galactosamine) gamowveul apoptozs, kaspaza

8-is inhibireba ufro gadamwyvetia am dros. ufro metic, dominant-

negatiuri FADD-is makodirebeli adenovirusi TNF/GalN-

inducirebuli apoptozis prevencias axdenda (Streetz et al., 2000). es

kvlevebi migvaniSneben im saintereso momentze, rom TNF–

inducirebul apoptozSi apoptozur kaskadSi Terapiuli

intervencia kaspaza 8-is doneze unda xdebodes. amasTan

dakavSirebiT, kaspaza 8-is endogenuri inhibitoris - “ujreduli

FADD-is msgavsi IL-1b-makonvertirebeli fermentis mainhibirebeli

proteinis” - eqspresiis gazrdaze orientirebuli Terapia Semdgom

yuradRebas imsaxurebs.

Fas-is mier gamowveuli apoptozis SeCereba SesaZlebelia

43

xsnadi Fas-iT genuri Terapiiis saSualebiT, rac potenciur

Terapiul gzas SeiZleba warmoadgendes. axlaxans aRweril iqna

sikvdilis receptoris mier gamowveuli apoptozis ori axali

inhibitori. suramins an neirokininis receptoris antagonistebs

SeuZliaT Tagvis modelebSi RviZlis toqsikuri dazianebis in vivo

inhibireba. suramini Sardovanas polisulfonirebul warmoebuls

warmoadgens. is axdens TNF-is, FasL-isa da qemoTerapiis Sedegad

gamowveuli apoptozis inhibirebas in vitro; in vivo naCvenebi iqna, rom

mas SeuZlia LPS-isa da D-galaqtozaminis erToblivi an Fas-is

agonisturi antisxeulis zemoqmedebiT gamowveuli masiuri

apoptozis Tavidan acileba. neirokinini-1 receptoris antagonistebi

Tagvebs Fas-isa da TNF-is mier gamowveuli RviZlis dazianebisgan

icaven. es SesaZlebeli xdeba antagonistebis mier ujredis

sikvdilis xelSemwyobi P naerTis moqmedebis antagonizirebiT.

1.4.3 mcenareuli warmoSobis nivTierebebi

mcenareebi sasargeblo wamlebisTvis kvlevis masalis

uzarmazar wyaros warmoadgenen. bolo wlebSi apoptozur procesze

zemoqmedebis unaris mqone mravali sxvadasxva mcenaris eqstraqti

iqna gamokvleuli. simsivnur ujredebSi aRniSnuli daavadebis

SesaCereblad programirebuli sikvdilis meqanizmebze zemoqmedebis

unari, romelic sxvadasxva mcenareul eqstraqts gaaCnia, intensiuri

da farTo kvlevebis sagania. Seswavlilia sxvadasxva ujreduli

xazi, rogoricaa HL60, adamianis hepatocelularuli karcinomis

ujreduli xazi, qolangiokarcinomis ujreduli xazi, B-ujredis

hibridomebi, U937 monocituri ujreduli xazi, HeLa ujredebi da

sxva. im agentebs Soris, romlebic ujredis programirebul

sikvdils iwvevdnen (rac izomeboda morfologiurad, an gamWoli

citofluorometris saSualebiT), aRmoCndnen iseTi mcenareebis

eqstraqtebi, rogoricaa fiTri da Semicarpus anacardium. dadginda

aseve, rom ujredis programirebul sikvdils da Sesabamisad

44

gamravlebis Sewyvetas iwvevdnen iseTi izolirebuli naerTebi,

rogorebicaa brionoluri mJava (Trichosanthes kirilowii var. Japonica-dan),

krocini (zafranadan) da alicini (nivridan). Cinuri mcenareuli

wamali “So-saiko-to” simsivnis garkveul ujredul xazebSi iwvevda

apoptozs. metad sainteresoa is faqti, rom JenSeni aferxebs Tmis

folikulebSi ujredis dasxivebiT gamowveul programirebul

sikvdils, rac mis mniSvnelovan Terapiul SesaZleblobebze

miuTiTebs. nutricevtuli saSualebebi (dieturi mcenarebi),

rogoricaa soio, niori, koWa, mwvane Cai da sxv., epidemiologiur

kvlevebSi miCneulia simsivnis SemTxvevebis Semamcirebel faqtorad.

maTi moqmedeba ukavSirdeba ujredis programirebuli sikvdilis

gamowvevis unars.

dadasturebuli iqna, rom sxvadasxva sakvebi da mcenareuli

warmoSobis wamlebi toqsiur efeqtebs iwvevs ujredis

programirebuli sikvdilis inducirebis gziT. magaliTad, virTxis

izolirebul hepatocitebSi ujredis programirebuli sikvdili

kavSirSi iyo diterpinoidebis Semcveli mcenareuli wamlis mier

gamowveul hepatitTan. sxva gamokvlevebis mixedviT, savaraudoa,

rom betel-sa da TambaqosTan dakavSirebuli piris “qerclovani”

karcinomebi SesaZloa asociirdebian p53–isa da c-myc–is

erToblivad moqmedebasTan, rac ujredis programirebuli

sikvdilis inhibirebas ganapirobebs. gamovlenilia mcenareebis mier

ujredis programirebuli sikvdilis modulaciis ramdenime

meqanizmi, maT Soris, endonukleazebis, p53-isa da kaspaza 3-is

aqtivacia (Bcl-2–damoukidebeli gziT), Tavisufali radikalebis

formirebis gaZliereba da sfinganinis akumulacia. ujredis

programirebuli sikvdili Tavdacvis metad konservirebuli

meqanizmia, romelic mcenareebsac gaaCniaT.

amrigad, ujredis programirebuli sikvdilis bunebriv

fenomenze zemoqmedeba misi modificirebis gziT SesaZloa

potenciurad sasargeblo wamlebs warmoadgendnen.

45

II Tavi - sakvlevi masala da meTodebi

eqsperimenti Catarda 20-25 gr masis mqone 50 zrdasrul

Tagvze. saeqsperimento cxovelebi erTnair pirobebSi imyofebodnen.

2.1 RviZlis imunuri dazianebis eqsperimentuli modeli

hepatitis eqsperimentuli modelis Sesaqmnelad 50 TagvSi

intraperitonealurad Seyvanili iqna bunebrivi mitogeni

konkanavalin–A (Con-A) 2mg/kg wonaze gadaangariSebiT.

eqsperimentuli cxovelebi davyaviT 3 jgufad. Con-A-s Seyvanidan 1

sT-is Semdeg cxovelebs erTxel kunTSi ukeTdeboda: 1. 0.2 ml

fiziologiuri xsnari (sakontrolo I jgufi), 2. plaferoni lb

(0.25mg/kg-ze) (II jgufi); 3. deqsametazoni _ 4 mg/kg-ze (III jgufi).

eqsperimentis dawyebidan 8, 24 da 48 saaTis Semdeg eTeriT

gabruebul Tagvebs vklavdiT cervikaluri dislokaciis meTodiT.

gamosakvlevad viRebdiT RviZls da sisxls. RviZls morfologiuri

kvlevisa da hepatocitebis funqciuri mdgomareobis Sesaswavlad,

sisxls ki - bioqimiuri gamokvlevisTvis.

2.2 histomorfologiuri meTodi

sinaTlis mikroskopuli kvlevisaTvis eqsperimentul masalas

(RviZli) fiqsaciisaTvis vaTavsebdiT formalinis 10%-ian xsnarSi,

vauwyloebdiT aRmavali koncentraciis spirtebSi da vayalibebdiT

parafinSi. rotaciul mikrotomze damzadebul 2-5 mkm sisqis

anaTlebs vRebavdiT hematoqsilin-eoziniT.

2.3 RviZlSi apoptozis Sefaseba imunohistoqimiuri

(TUNEL)LmeTodiT

hepatocitebis apoptozi fasdeboda dnm-is fragmentaciiT

terminaluri deoqsinukleotidil transferazaze-damokidebuli

46

(tdt) dUTP-biotinis (TUNEL) moniSvnis meTodis gamoyenebiT. yvela

safexuri xorcieldeboda TandarTuli protokolis mixedviT.

amokveTili RviZli fiqsirdeboda 10%-ian formalinSi da

yalibdeboda parafinSi. tardeboda parafinSi Cayalibebuli,

rotaciul mikrotomze daWrili 5-mikroniani anaTlebis

deparafinizacia da rehidratacia, Semdeg inkubacia proteinaza K–

Ti (20mg/ml 100mM da 50 mM edta) 15 wuTi 25oC-ze. Semdeg yvela

anaTali irecxeboda distilirebuli wyliT, anaTlebi

inkubirdeboda buferSi 5wT. Semdeg emateboda tdt da

biotinmoniSnuli deoqsinukleotidebis Tanaobisas tenian yuTSi

midinareobda inkubacia 1 saaTiT 37C-ze. reaqciis Sewyveta xdeboda

gamrecxi buferis saSualebiT. moniSnuli slaidebis daTvaliereba

xdeboda sinaTlis mikroskopSi. qsovilebi iRebeboda hematoqsilin-

eozinSi.

2.4Aazotis oqsidisa da glutaTion reduqtazas atiurobis

gansazRvra

sisxlis SratSi azotis oqsidis raodenoba fasdeboda

arapirdapiri meTodiT grisis reaqciis gamoyenebiT, romelic

warmoqmnis qromofors nitritTan reaqciis Sedegad. Sratis 100ml-s

vainkubirebdiT 100ml grisis reaqtivTan 96-fosoian plataSi.

inkubacia mimdinareobda 10wT-is ganmavlobaSi oTaxis

temperaturaze. nitritis koncentracia fasdeboda

speqtrofotometrulad multiskanze (Multiscan, Lab Systems, fineTi).

absorbcia isazRvreboda 540nm talRis sigrZeze. yvela nimuSi

keTdeboda tripletebSi.

glutaTionreduqtazas aqtiurobis gansazRvra xdeboda

glutaTion reduqtazas nakrebis saSualebiT (Sigma-Aldrich, USA).

aqtiuroba izomeboda absorbciis zrdis cvlilebiT DTNB [5,5’-

47

dithiobis(2-nitrobenzoic acid)] substratis damatebisas 412nm talRis

sigrZeze.

2.5 hepatocitebis gamoyofa da maTi mitoqondriuli

aqtiurobis Seswavla tetrazoliumze dafuZnebuli

kolorimetriuli MTT testiT

RviZlis homogenizaciisa da garecxvis Semdeg vamzadebdiT

hepatocitebis kulturas ujredebis koncentraciiT – 1 mln/ml.

ujredTa suspenzias TiTo ml-is raodenobiT vaTavsebdiT

sinjarebSi. Semdeg hepatocitebis suspenzias 150 mkl raodenobiT

vasxamdiT 96-fosoian brtyelZirian mikroplanSetis fosoebSi (Libro

Plastics).

eqsperimentis yovel variantSi cda da kontroli

sruldeboda 3 fosoSi. Semdeg mikroplanSetis inkubirebas

vaxdendiT TermostatSi 100% tenianobis, haerSi naxSirorJangis 5%-

iani Semcvelobisa da 370C temperaturis pirobebSi 4 saaTis

ganmavlobaSi.

vamzadebT dimeTilTiazolil difenil tetrazolium

bromidis (MTT Sigma) xsnars - 5 mg reaqtivi 1ml fosfaturi buferis

marilxsnarze.

TiToeul fosos vumatebdiT 10-10mkl MTT-is momzadebul

xsnars.

Semdeg planSets sainkubaciod vdgamdiT 1sT-iT 370C

temperaturaze СO2 inkubatorSi. inkubaciis damTavrebisas planSets

vamowmebdiT mikroskopis qveS. formazanis kristalebis warmoqmnis

Semdeg, supernatants frTxilad aviwovdiT da TiToeul fosoSi

vumatebdiT 100-100mkl 10%-ian natriumdodecilsulfats.

vaikubirebdiT 2sT-iT 370temperaturaze, CO2 inkubatorSi.

Sedegebis wakiTxvas vaxdendiT multiskanis saSualebiT 540-

570nm talRis sigrZeze.

48

2.6 hepatocitebSi kaspazebis aqtiurobis gansazRvra

kaspaza 3 da kaspaza 8-s aqtiurobis gansazRvra xdeboda

kolorimetruli nakrebis saSualebiT. mwarmoeblis instruqciis

mixedviT, hepatocitebis suspenzias vumatebdiT lizis bufers.

Semdeg emateboda kaspaza-3-is (Ac-DEVD) an 8-s (Ac-IETD) substrats,

romelic dakavSirebuli iyo p-nitroanilinTan da vainkubirebdiT

37°C–ze 90 wuTiT. speqtrofotometrze (Multiscan, Labsystems, Finland)

vsazRvravdiT optikur simkvrives 405nm talRis sigrZeze.

gamoyofili nitroanilinis raodenoba homogenatSi kaspazebis

aqtiurobis pirdapirproporciulia. optikuri simkvrivis gazomva

xdeboda yovel 5 wuTSi da gamoTvlili iqna Sesabamisad saSualo

da standartuli gadaxra.

2.7 ALT-s gansazRvris speqtrometriuli meTodi

sisxlSi alaninaminotransferazas (ALT) aqtivobas

vswavlobdiT speqtrofotometriuli meTodiT. amisaTvis Tagvidan

viRebdiT sisxls, vacentrifugirebdiT 3000 g-ze 15 wT-is

ganmavlobaSi, ris Semdegac miRebul sisxlis Srats (raodenobiT

0.05 ml) vumatebdiT 0.25 ml ALT-s standartul substrats

(fosfaturi buferi 0.1 moli/l, Semdeg 0.2 moli/l DL--alanins da

2 mmoli/l 2-oqsiglutarats). amis Semdeg vaxdendiT sinjis

inkubirebas 370C temperaturaze 60 wT-is ganmavlobaSi, vumatebdiT

0.25 ml 2-4-dinitrofenilhidrazinis xsnars (1 mmoli/l, HCI 1

mmoli/l). miRebul sinjs vurevdiT da vtovebdiT 20 wT-is

ganmavlobaSi 220C temperaturaze. amis Semdeg vumatebdiT 2.5 ml

NaOH-is xsnars, vtovebdiT 10 wT-is ganmavlobaSi da optikur

simkvrives vzomavdiT 530 nm talRis sigrZeze sakontrolo xsnaris

mimarT, sadac sisxlis Sratis nacvlad damatebuli iyo 0.25 ml

fiziologiuri xsnari.

49

ALT-s koncentracias vsazRvravdiT standartuli grafikis

saSualebiT.

2.8 masalis statistikuri damuSaveba

masala damuSavda statistikuri programis Statistica 6.0 (Statsoft,

Mineapolis, USA) gamoyenebiT.

50

III Tavi - sakuTari gamokvlevis Sedegebi

3.1 plaferoni lb-s zegavlena Con-A inducirebuli msubuqi

hepatitis dros sisxlSi alaninaminotransferazas

Semcvelobasa da RviZlSi ganviTarebul morfologiur

cvlilebebze

diagrama 1-ze mocemulia Con-A inducirebuli hepatitis dros

Tagvebis sisxlSi ALT-s cvlilebebi dakvirvebis sxvadasxva vadaze

(normaSi aRniSnuli maCvenebeli udris 0.5-s).

diagrama #1

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

Before injection 8 24 48

Con-A

ConA+PL

ConA+Dex

dakvirvebis I vadaze (8 sT) ALT-s done sakontrolo Tagvebis

sisxlSi TiTqmis 3-jer aRemateba normas. rac Seexeba

deqsametazoniT da plaferoniT namkurnaleb Tagvebs, maT sisxlSi

51

ALT-s done orive SemTxvevaSi erTnairia da TiTqmis 2-jer

aRemateba normas.

dakvirvebis II vadaze (24 sT) sakontrolo da namkurnalebi

Tagvebis sisxlSi ALT-s done TiTqmis erTnairia da 2-jer aRemateba

normas.

dakvirvebis III vadaze (48 sT) sakontrolo Tagvebis sisxlSi

ALT-s done kvlav 2-jer aRemateba normas, deqsametazoniT

namkurnalebi Tagvebis sisxlSi ki ALT-s done dakvirvebis am

droisaTvis kvlav statistikurad sarwmunod maRalia normasTan

SedarebiT, plaferoni lb-s zemoqmedebiT ki aRniSnuli maCvenebeli

normas ubrundeba.

aRniSnulidan gamomdinare, deqsametazoni dakvirvebis I

vadaze amcirebs ALT-s dones, xolo, Semdgom vadebze igi

statistikurad sarwmunod aRar gansxvavdeba sakontrolo Tagvebis

analogiuri maCveneblisagan. plaferoni lb-s zegavleniT

dakvirvebis III vadaze ALT done normas ubrundeba.

.

sur. 1 janmrTeli Tagvis RviZli morfologiuri preparati

52

Con-A-s ineqciidan 8 sT-is Semdeg Tagvebis RviZlSi mwvave

anTebisaTvis damaxasiaTebeli morfologiuri cvlilebebi

aRiniSneba. hepatocitebis didi nawili vakuolur distrofias

ganicdis. amasTan erTad, fokaluri nekrozis kerebic gvxvdeba.

nekrozuli hepatocitebis citoplazma mkveTrad eozinofiluria,

rac nekrozis ubnebSi eozinofiluri (e.w. kaunsilmanis)

sxeulakebis saxiT vlindeba. alag-alag orbirTviani hepatocitebis

arseboba aRiniSneba (sur. 2).

sur. 2 sakontrolo jgufi ConA-s ineqciidan 8 sT-is Semdeg

RviZlis parenqimaSi limfoiduri rigis ujredebis

infiltratebi gvxvdeba. infiltracia gansakuTrebiT intensiuria

portul sivrceebSi. damaxasiaTebelia centraluri venebisa da

RviZlis sinusoidebis gaganiereba. gamoxatulia “slaj-fenomeni”.

53

sur. 3 ConA-s ineqciidan 24 sT-is Semdeg

Con-A-s Seyvanidan 24 sT-is Semdeg aRniSnuli cvlilebebi

kvlav intensiuradaa gamoxatuli. hepatocitebis umetesoba

vakuolur distrofiasa da nekrozul cvlilebebs ganicdis.

ujredebis umravlesoba hgavs erT did vakuols (sur. 3)

msgavsi suraTia eqsperimentis dawyebidan 48 sT-is Semdegac.

wina vadebTan SedarebiT aRiniSneba orbirTviani hepatocitebis

raodenobis mateba, rac regeneraciuli procesebis gaaqtivebaze

miuTiTebs. zogierT areSi RviZlis normaluri trabekuluri

struqtura vlindeba. imave vadaze zogierTi normaluri

hepatocitic Cndeba (sur. 4).

54

sur. 4 ConA-s ineqciidan 48 sT-is Semdeg

Con-A-sTan erTad deqsametazonis Seyvanis fonze zemoT

aRniSnuli cvlilebebi SedarebiT naklebi intensivobiT xasiaTdeba.

eqsperimentis dawyebidan 8 sT-is Semdeg Tagvebis RviZlSi

hepatocitebis didi nawili vakuolur distrofias ganicdis, Tumca

wilakebis ZiriTadi trabekuluri struqtura SenarCunebulia.

distrofiuli cvlilebebis fonze aRiniSneba mcire zomis

nekrozuli ubnebis arseboba. kaunsilmanis sxeulakebi upiratesad

periportul zonebSi gvxvdeba (sur. 5).

sur. 5 ConA-s ineqciidan 8 sT-is Semdeg, deqsametazoniT namkurnalevi

55

portuli sivrceebis limfoiduri infiltracia umniSvneloa.

“slaj-fenomeni” sustad aris gamoxatuli. umetes SemTxvevaSi

eriTrocitebis konturebi Senaxulia. aqa-iq gvxvdeba mcire

raodenobiT orbirTviani hepatocitebi.

24 sT-is Semdeg RviZlSi mniSvnelovnad matulobs

orbirTviani hepatocitebis raodenoba. amave dros aRiniSneba

destruqciuli cvlilebebis gaRrmaveba .

sur. 6 ConA-s ineqciidan 24 sT-is Semdeg, deqsametazoniT

namkurnalevi

limfoiduri infiltraciis arseboba damaxasiaTebeli ar

aris. 48 sT-is Semdeg destruqciuli cvlilebebi naklebad aris

gamoxatuli, Tumca, hepatocitebis nawilSi kvlav vakuoluri

distrofiisaTvis damaxasiaTebeli cvlilebebi aRiniSneba.

nekrozuli ubnebis arseboba iSviaTia da mniSvnelovnad aris

momatebuli orbirTviani hepatocitebis raodenoba. limfoiduri

infiltratebis arseboba damaxasiaTebeli ar aris (sur. 7).

56

sur. 7 ConA-s ineqciidan 48 sT-is Semdeg, deqsametazoniT

namkurnalevi

Con-A-sTan erTad plaferoni lb-s Seyvanis SemTxvevaSic

eqsperimentis dawyebidan 8 sT-is Semdeg RviZlSi mwvave

anTebisaTvis damaxasiaTebeli morfologiuli suraTia.

hepatocitebis didi umravlesoba vakuolur distrofias ganicdis.

amasTanave, aRsaniSnavia, rom kaunsilmanis sxeulakebis raodenoba

umniSvneloa, xolo wilakis trabekuluri Seneba SenarCunebulia

(sur. 8). aseve iSviaTi da umniSvneloa limfoiduri infiltraciis

ubnebi.

sur. 8 ConA-s ineqciidan 8 sT-is Semdeg, plaferoniT namkurnalevi

57

24 sT-Tvis hepatocitebis umravlesoba kvlav distrofiul

cvlilebebs ganicdis, magram nekrozuli ubnebis arseboba

damaxasiaTebeli ar aris (sur. 9).


48 sT-isaTvis wilakis normuli Seneba ZiriTadad

aRdgenilia, Tumca, hepatocitebis nawilSi kvlav vakuoluri

distrofiis niSnebi aRiniSneba. nekrozuli ubnebisa da

limfoiduri infiltratebis arseboba ar aRiniSneba (sur. 10).


58

amgvarad, Con-A inducirebuli hepatitis dros plaferoni lb,

deqsametazonis msgavsad, RviZlis qsovils Rrma destruqciuli

cvlilebebis ganviTarebisagan icavs da RviZlis wilakis normuli

Senebis aRdgenas aCqarebs.

59

3.2 Con-A inducirebuli msubuqi hepatitis dros RviZlsa da

sisxlSi mimdinare oqsidaciuri procesebis maCveneblebis

modulacia imunomodulatorebiT

reaqtiuli Jangbaduri saxeobebis (ROS) ujredSida

koncentracia mkacrad kontrolirdeba dacvis mravalricxovani

meqanizmebiT, maT Soris, ROS-is skavenjeri fermentebisa da

patara antioqsidanti molekulebis mier. am antioqsidanti

sistemebidan aRsaniSnavia glutaTioni da masze damokidebuli

fermentebi, romlebic oqsidaciuri dazianebisagan damcav

meqanizms warmoadgenen rogorc cirkulaciaSi, ise sxvadasxva

qsovilSi, maT Soris, RviZlSi (Gate et al, 1999). glutaTionis

redoqs stratusis Secvla mravalgvar mJangavi molekulas

SeuZlia. es statusi, Cveulebriv, uzrunvelyofilia glutaTionis

damSleli da aRmdgeni fermentebis mier (Halliwell et al, 1996; Hayes et

al., 1999). winamdebare kvlevaSi Cven ganvsazRvreT azotis oqsidis

koncentraciebi da glutaTion reduqtazas aqtiuroba SratSi

msubuqi hepatitis eqsperimentul modelSi. garda amisa, Cven

SevafaseT izolirebuli hepatocitebis mitoqondriuli

aqtiuroba.

Con-A inducirebidan 8sT-is Semdeg Tavisufali azotis

oqsidis raodenoba plazmaSi matulobs jgufis sakvlev

cxovelebSi daaxloebiT 2-jer izrdeba. II da III jgufebSic misi

sagrZnoblad zrda SeiniSneba, rac TiTqmis 2-jer aRemateba

janmrTeli Tagvis monacems. Con-A inducirebidan 24sT-is Semdeg

Tavisufali azotis oqsidis epr signalis intensivoba I jgufis

cxovelebSi normasTan SedarebiT kvlav maRalia. II jgufis

cxovelebSi NO-s epr signalis intensivoba wina vadasTan

SedarebiT statistikurad mniSvnelovnad ar icvleba da kvlav

momatebulia. III jgufis cxovelebSi NO-s epr signalis

intensivoba normasTan SedarebiT kvlav maRalia, magram wina

60

vadis monacemTan SedarebiT mcirdeba (p<0,01) da sakontrolo

maCvenebelTan SedarebiT 43%-iT naklebia. Con-A inducirebidan

48sT-is Semdeg Tavisufali azotis oqsidis (NO) epr signalis

intensivoba I jgufis cxovelebSi normasTan SedarebiT kvlav

maRali rCeba da wina vadis monacemTan SedarebiT umniSvnelod

mcirdeba. aRniSnuli maCvenebeli II da III jgufis cxovelebSic

isev sagrZnoblad maRalia da wina vadis monacemebis Sesabamisad

72%-s da 140%-s (p<0,01) Seadgens.

plaferoni lb da deqsametazoni xels uwyoben NO-s maRali

donis SenarCunebas, amasTan, plaferoni lb mis cvlilebas

axdens.

Con-A-Ti zemoqmedebidan 8sT-is Semdeg NO-is

koncentraciebi momatebulia da ar xasiaTdeba mniSvnelovani

cvlilebebiT eqsperimentis bolomde. I da III jgufebTan

SedarebiT plaferoni lb–Ti zemoqmedeba mniSvnelovnad amcirebs

NO-is koncentraciebs 24-e saaTze. Tumca, plaferoni lb-is

zegavleniT azotis oqsidis sinTezi mniSvnelovnad gaZlierda

48sT-is Semdeg Con-A–is ineqciis gamo (diagrama 2).

diagrama 2 SratSi nitrit/nitratis doneebis cvlilebebi Con-A-inducirebul eqsp. msubuq hepatitSi

0.250 Before injection Con-ACon-A+PLB Con-A+DEX

0.200

Op

tica

l den

sity

0.150

0.100

0.050

0.000 8 24 48

61

garda amisa, Con-A–Ti damuSavebul TagvebSi RviZlis

dazianeba intravenuli ineqciidan 8sT-is Semdeg plazmuri

transaminazebis momatebuli koncentraciebiT xasiaTdeba, rac

piks 24-e sT-ze aRwevs.

Cven SeviswavleT antioqsidanturi indikatori, kerZod,

glutaTion reduqtaza. gamokvlevis Sedegebma gviCvena, rom me-8

sT-ze glutaTion reduqtazas aqtiuroba I jgufSi normalur

cxovelebTan SedarebiT gazrdili iyo (p<0,01) (diagrama 3).

diagrama 3 SratSi glutaTion reduqtazas aqtiuroba eqsp. msubuqi hepatitis dros

0.070 Before injection Con-A

0.060 Con-A+PLB Con-A+DEX

0.050

0.040

0.030

0.020

0.010

0.000 8 24 48

deqsametazoniTYzemoqmedebiT, glutaTion reduqtazas done

I vadaze sagrZnobad izrdeba, Semdeg vadebze ki mcirdeba, magram

normas ar ubrundeba. plaferoni lb–iT zemoqmedebas I jgufTan

SedarebiT glutaTion reduqtazas aqtiurobis mniSvnelovani

Semcireba mosdevda ise, rom Semcirebuli done aRar icvleboda

Semdgomi 48sT-is ganmavlobaSi.

Cven mier gamovlenili monacemebi Seesabameba sxva kvlevis

Sedegebs, romlis mixedviTac, cirozsa da hepatocelularul

karcinomaSi sakontrolo qsovilTan SedarebiT glutaTion

62

reduqtazas aqtiuroba gazrdilia (Czeczot et al., 2006). ujredis

doneze glutaTionis homeostazi uzrunvelyofilia biosinTezs,

moxmarebas, oqsidaciasa da eqsports Soris balansiT.

homeostazis darRvevam ki SeiZleba imoqmedos RviZlis unarze

uzrunvelyos oqsidaciuri dazianebisgan dacva (Fernandez-Checa et

al., 1997).

ufro metic, MTT meTodis saSualebiT SevafaseT

hepatocitebis mitoqondriuli funqcia da imunomamodulirebeli

wamlebis efeqti. hepatocitebis mitoqondriuli funqcionireba

sakontrolo cxovelebTan SedarebiT gaZlierda I jgufSi.

plaferoni lb mniSvnelovnad zrdis mitoqondriul aqtiurobas

Con-A-Ti zemoqmedebidan 48sT-is Semdeg. deqsametazons igive

efeqti moaqvs 24-e saaTze. es Sedegebi adastureben Cven mier

manamde miRebul monacemebs da miuTiTeben imaze, rom plaferoni

aZlierebs mitoqondriul funqcias da xels uwyobs

regeneraciul process RviZlSi.

diagrama 4 hepatocitebis spontanuri stimulaciis indeqsi MTT testis saSualebiT

0.700 Before inje tionc Con-AConA+PLB Con-A+DEX

0.600

0.500

0.400

0.300

0.200

0.100

0.000 24h 48h

saboloo jamSi Cven movaxdineT imis demonstrireba, rom

oqsidanti/antioqsidantis sistema eqsperimentuli msubuqi

63

hepatitis dros balans kargavs. es faqti eTanxmena hipoTezas,

romlis mixedviTac, oqsidaciuri stresi RviZlis

zemoTaRniSnuli daavadebebis ganviTarevaSi mniSvnelovan rols

TamaSobs. ismis mTavari kiTxva: es darRveva RviZlis mocemuli

daavadebebis erT-erTi gamomwvevi mizezia Tu maTi ganviTarebis

mxolod erT-erTi Sedegi? saWiroa Semdgomi kvleva aRniSnul

kiTxvaze pasuxis gasacemad. qsovilebidan reaqtiuli Jangbaduri

saxeobebisa da Tavisufali radikalebis mocilebis daqveiTebuli

unari erT-erTi faqtoria, romelmac SeiZleba neoplaziis

ganviTareba ganapirobos.

64

3.3 Con-A inducirebuli msubuqi hepatitis dros RviZlSi

kaspazebis aqtiuroba da maTi modulacia

imunomodulatorebiT

kaspazebis gaaqtiureba apoptozuri procesis damaxasiaTebeli

niSania. apikaluri kaspazebis regulacia SeiZleba gadamwyveti

faqtori iyos am procesSi. kaspaza 8-is TviTgaaqtiureba iwvevs

kaspaza 3-is pirdapir gaaqtiurebasa da Bid–is gaxleCvas (Li et al.,

1998).

winamdebare kvlevis mTavari miznebi iyo: 1. imis dadgena, Tu

romeli kaspazebis gaaqtiureba xdeboda in vivo Con-A–s

administrirebis Sedegad; 2. imis dadgena, Tu rogori efeqtebi

hqonda Sesabamisad plaferoni lb–sa da deqsametazons am

gaaqtiurebaze. kerZod, kolorimetruli meTodiT SeviswavleT zeda

kaspaza-8 da qveda kaspaza-3. gamovikvlieT, agreTve, SratSi ALT–is

koncentraciebi.

mwvave eqsperimentuli hepatitisgan gansxvavebiT (SaraSeniZe,

2003), Cveni kvlevis Sedegebma aCvenes, rom ALT–is koncentraciebi

Con-A–s administrirebidan 8sT-is Semdeg momatebuli iyo da

maqsimums 48-e sT-ze aRwevda. Con-A jgufTan SedarebiT,

deqsametazonis gamoyeneba ar cvlida ALT–is koncentraciebs.

plaferoni lb–Ti zemoqmedebis SemTxvevaSi ki ALT–is

koncentraciebma I jgufze ufro metad moimates (diagrama 1).

arsebuli kvlevevis Sedegebi, romlebmac Con-A–s

administrirebis Sedegad prokaspaza-8-isa da prokaspaza-3-is

procesingis demonstrireba moaxdines, mkafiod Seesabameba Cvens

mier miRebul monacemebs. kerZod, vaCveneT, rom Con-A–Ti

zemoqmedebidan me-8, 24-e da 48-e sT-ze kaspaza-8-isa da kaspaza-3-is

aqtiuroba aSkarad saxeze iyo. am kaspazebis aqtiurobebis maqsimumi

modioda me-8 sT-ze, 24-48 sT-ebSi ki gamudmebiT mcirdeboda, Tumca,

48-e sT-ze normaSi mainc ar iyo.

65

amis Semdeg Cven davadgineT, rom plaferoni lb-s me-8 da 24-e

sT-ebze kaspaza-3-isa da kaspaza-8-is aqtiurobis gaZlierebis

tendencia gaaCnia. Tumca, es cvlilebebi mniSvnelovani ar aris.

plaferoni lb orive kaspazis aqtiurobas arsebiTad mxolod 48sT-

isTvis zrdida (p<0,01). Cven mier Catarebuli analizis Sedegebis

mixedviT, deqsametazoni Cven mier Seswavlil nebismier etapze

axdenda orive kaspazis inhibirebas (diagramebi 5, 6).

diagrama 5 kaspaza-3-is aqtiuroba Con-A–inducirebuli hepatitis dros

0.350 Control ConA ConA+PLB 0.300 ConA+DEX

0.250

0.200

0.150

0.100

0.050

0.000 8h 24h 48h

kaspasa-8-is aqtiuroba Con-A inducirebuli msubuqi hepatitis dros diagrama 6 kaspaza-8-is aqtiuroba Con-A–inducirebuli

hepatitis dros

0.000

0.050

0.100

0.150

0.200

0.250

0.300

0.350

8h 24h 48h

ControlCon-A

Con-A+PLBCon-A+DEX

66

aRsaniSnavia, rom wina kvlevebis mier naCvenebi iqna

plaferoni lb–s antiapoptozuri efeqti mwvave hepatitis

modelebSi da klinikur gamocdebSi (gongaZe, 2004). Cven ki

davadgineT, rom plaferoni lb proapoptozur Tvisebasac avlens,

rac gamoixateba msubuqi hepatitis dros kaspaza-3-isa da kaspaza-8-

is aqtiurobis gazrdaSi.

naCvenebia, rom Warb apoptozs adgili aqvs mwvave da

qronikuli virusuli hepatitis, alkoholuri da araalkoholuri

hepatitis, RviZlis qolestaturi daavadebis, vilsonis daavadebisa

da “implantanti maspinZlis winaaRmdeg daavadebaSi” (GVHD). mudmivi

apoptozi ukavSirdeba RviZlis fibrozis ganviTarebas. meore mxriv,

arasakmarisi apoptozi asociirebulia RviZlSi da sanaRvle

sistemaSi simsivnis ganviTarebasa da progresiasTan. ufro metic,

interferonis antivirusuli efeqti mediirdeba apoptozis

gamowvevis gziT (Castelli et al, 1997).

amrigad, Cveni kvleva – Cven wina kvlevebTan erTad –

gvavaraudebinebs, rom plaferoni lb–s RviZlis sxvadasxva

dazianebis, maT Soris, hepatitisa da simsivnebis Terapiuli

potenciali gaaCnia. cxadia, saWiroa Semdgomi klinikuri

gamokvlevebi am rolSi plaferoni lb–s efeqturobis srulad

Sesaswavlad.

67

3.4 eqsperimentuli msubuqi hepatitis dros hepatocitebis

apoptozuri cvlilebis Seswavla imunohistoqimiuri meTodiT

TUNEL-is meTodis gamoyenebiT SeviswavleT eqsperimentuli

msubuqi hepatitis dros RviZlSi mimdinare apoptozuri

cvlilebebi. eqsperimentuli kvlevis Sedegad miRebuli monacemebi

warmodgenilia cxrili #1-Si.

cxrili #1 apoptozuri birTvebis raodenobrivi maCveneblebi Tagvebis hepatocitebSi Con-A-Ti gamowveuli eqsperimentuli msubuqi hepatitis

dros

vada / sakvlevi jgufebi

janmrTeli konkanavalin-A konkanavalin-A + plb

konkanavalin-A + deqsametazoni

8 saaTi 54,23 ± 9,09 154,39±15,22 180,39±17,2 164,39±15,02

24 saaTi “ – “ 76,41±12,12 82,34±11,15 86,14±9,11

48 saaTi “ – “ 75,41±10,11 280,23± 15,21 84,12±9,10

rogorc cxrilidan Cans, janmrTeli Tagvebis hepatocitebSi

yovel 10000 ujredze saSualod 54,23±9,09 apoptozuri birTvi

aRiniSneboda. damaxasiaTebeli iyo mcire zomis, baci yavisferi

apoptozuri birTvebis Tanabari ganawileba.

janmrTel TagvebTan SedarebiT, eqsperimentis I vadaze (8 sT)

apoptozuri birTvebis raodenoba mniSvnelovnad matulobs (p<0,01)

da yovel 10 000 ujredze saSualod 154,39±15,22 apoptozuri birTvi

aRiniSneba (cxrili #1). plaferoni lb-Ti da deqsametazoniT

mkurnaloba apoptozze gavlenas ar axdens, razec apoptozuri

birTvebis umniSvnelo cvlileba miuTiTebs (P>0,05).

24 sT-ze yvela jgufSi apoptozuri birTvebis raodenoba I

vadasTan SedarebiT statistikurad sarwmunod mcirdeba (P<0,05) da

jgufebs Soris sxvaoba ar aRiniSneba (P>0,05).

eqsperimentis bolo vadaze (48 sT) sakontrolo da

deqsametazoniT namkurnalebi jgufebis cxovelebis RviZlSi

68

apoptozuri birTvebis raodenoba TiTqmis igivea, rac 24 sT-ze iyo

(suraTi #11 da suraTi #12). plaferoni lb-Ti namkurnaleb

TagvebSi ki am vadaze apoptozuri birTvebis raodenoba daaxloebiT

2-jer matulobs (sur. #13).

suraTi #11 sakontrolo jgufis Tagvis RviZli SeRebili TUNEL-is meTodi, 48 saaTi

suraTi #12 deqsametazoniT namkurnalebi Tagvis RviZli SeRebili TUNEL-is meTodiT, 48 saaTi (apoptozuri birTvebi miTiTebulia isrebiT).

69

suraTi #13 plaferoni lb-Ti namkurnalebi Tagvis RviZli SeRebili TUNEL-is meTodiT, 48 saaTi (apoptozuri birTvebi miTiTebulia isrebiT).

amrigad, Cvens mier Catarebuli kvlevis Sedegad gamovlinda,

rom eqsperimentuli msubuqi hepatitis modelSi plaferoni lb-s

gamoyeneba 48 sT-ze RviZlSi sarwmunod zrdis apoptozuri

hepatocitebis raodenobas, rac metyvelebs am preparatis

proapoptozur efeqtze. maSin, roca deqsametazoni aseT efeqts ar

avlens.

70

IV Tavi – sakuTari Sedegebis ganxilva

virusuli hepatitebi hepatologiis umniSvnelovanes problemas

warmoadgens. farTo gavrcelebis, mZimed garTulebebis, maRali

letalobisa da qronizaciis gamo B (HBV) da C (HCV) hepatitebs

gansakuTrebuli mniSvneloba eniWebaT. yovelwliurad B hepatitisgan 2

mln adamiani iRupeba. jandacvis msoflio organizaciis monacemebiT, 10-

20 weliwadSi qronikuli C hepatitiT gamowveuli RviZlis ciroziTa da

hepatocelularuli kiboTi daavadebulTa ricxvi daaxloebiT^60-68%-iT

moimatebs. miuxedavad imisa, rom virusuli hepatitebis paTogenezSi

ujred-damokidebuli imunuri pasuxi mniSvnelovan rols TamaSobs,

faqtorebi romlebic hepatitebis qronizacias ganapirobebs dRemde

ucnobia.

cnobilia, rom orive – hepatiti B da C - virusi

hepatotropulia, Tumca isini pirdapiri citopaTiurobiT ar

gamoirCeva (Lok et al., 2001; Liang et al., 2000). epidemiologiuri kvlevebis

mixedviT, HHCV virusiT inficirebulebis 80-85%-Si ver xerxdeba virusis

sruli eliminacia, rac SemdgomSi qronikuli hepatitis ganviTarebas

uwyobs xels (Liang et al., 2000; Lauer et al., 2001). hepatocitebis lizisi

organizmis imunuri pasuxis siZlierezea damokidebuli (Dudley et al., 1972).

savaraudoa, rom am mxriv wamyvani roli hepatocitebis apoptozs eniWeba.

RviZlis mwvave dazianebisas hepatocitebis apoptozis siWarbe RviZlis

funqciuri ukmarisobisa da komis safuZveli xdeba (Gremion et al., 2004;

Kountouras et al., 2003), misi deficiti ki avTvisebiani transformaciis,

virusuli latenturi mimdinareobisa da autoimunuri daavadebebis

ganviTarebas ganapirobebs (Solary et al., 1996). inficirebuli hepatocitebis

apoptozis saSualebiT organizmi virusuli infeqciis blokirebasa da

misgan Tavdacvas axerxebs. virusuli genomi akodirebs cilebs,

romlebic apoptozuri procesis daTrgunvas iwvevs da amiT Tavs aRwevs

imunuri pasuxisgan. ase magaliTad HCV virusis core proteini

apoptozuri procesis regulacias axdens misi gaZlierebis an

inhibirebis gziT.

71

iseTi farmakologiuri nivTierebebis gamovlena, romlebic

apoptozis daTrgunvas an stimulacias, agreTve I tipis citokinebis

produqcia/aqtivobas aregulireben da/an II tipis citokinebis moqmedebas

astimulireben, RviZlis imunuri dazianebis profilaqtikisa da

mkurnalobisaTvis potenciur kandidatebs warmoadgenen (Okamoto et al.,1998;

Hershkoviz et al.,1999; Okamoto, Kanda 1999; Okamoto et al., 2000).

plaferoni lb adamianis placentidan miRebuli peptidebis

narevia. misi efeqti HBV infeqciis mqone pacientebis HBs antigeniT

stimulirebeli limfocitebis proliferaciul aqtivobaze daavadebis

fazazea damokidebuli. daavadebis mwvave fazaSi igi aRniSnul

maCvenebels Trgunavs, xolo qronikul fazaSi ki zrdis. plaferoni lb

gavlenas axdens citokinebis: interleikin-1-is, interferon gamas,

simsivnis nekrozis faqtori-alfasa da interleikin-4-is sinTezze

(Chikovani et al., 1999). yovelive zemoTTqmulma gvafiqrebina, rom plaferoni

lb RviZlis msubuqi imunuri dazianebis SemTxvevaSi efeqturi unda

yofiliyo.

konkanavalin A-Ti (Con A) gamowveuli RviZlis dazianeba

autoimunuri hepatitis farTod gavrcelebul eqsperimentul models

warmoadgens (Tiegs et al., 1992; Louis et al., 1997; Kato et al., 2001). konkanavalin A-s

zemoqmedebiT RviZlSi CD4+ T limfocitebi grovdeba da FasL-is eqspresia

izrdeba, romelic, Tavis mxriv, citotoqsiurobis ganviTarebas

ganapirobebs. garda amisa, Con A-inducirebuli hepatitis ganviTarebaSi

gadamwyvet rols asruleben TH1/TH2 tipis citokinebi, rogorebicaa:

interleikin 1 (IL-1), IL-12, IL-4, IL-10, (Ksontini et al., 1998; Tiegs et al, 1997; Gantner

et al, 1995; Trautwein et al., 1998; Kimura et al., 1999). RviZlis msubuqi dazianebis

dros apoptozis rolis Sesaswavlad Con A-inducirebuli msubuqi

hepatitis eqsperimentuli modeli gamoviyeneT. amave modelze

SeviswavleT apoptozis mimdinareobaze preparatebis – plaferoni lb-

s da deqsametazonis – gavlena.

72

Con-A-s Seyvanidan 1 sT-is Semdeg Tagvebis kunTSi Segvyavda

plaferoni lb, fiziologiuri xsnari an deqsametazoni. dakvirvebas

vaxdendiT eqsperimentis 3 vadaze: hepatitis inducirebidan 8, 24 da 48

saaTis Semdeg. aRniSnul vadebze aRebul masalas (RviZli, sisxli)

bioqimiuri, imunohistoqimiuri da imunologiuri meTodebiT

vikvlevdiT.

rogorc gamokvlevis Sedegebis analizma cxadyo, dakvirvebis I

vadaze (8 sT) ALT-s done sakontrolo Tagvebis sisxlSi matulobs

(diagrama 1), rac Seexeba deqsametazoniT da plaferoniT namkurnaleb

Tagvebs, maT sisxlSi ALT-s done orive SemTxvevaSi erTnairia da igi

sakontrolo jgufis cxovelebis analogiur monacemTan SedarebiT

statistikurad sarwmunod ar gansxvavdeba.

dakvirvebis II vadaze (24 sT) sakontrolo da namkurnalebi

Tagvebis sisxlSi ALT-s done matulobs I vadis Sesabamis monacemebTan

SedarebiT da amasTan erTmaneTisgan statistikurad sarwmunod ar

gansxvavdeba. dakvirvebis III vadaze (48 sT) sakontrolo Tagvebis

sisxlSi ALT-s donis matebis tendencia SenarCunebulia,

deqsametazoniT namkurnaleb TagvebSi aRiniSneba klebis tendencia,

xolo plaferoni lb-s zemoqmedebiT aRniSnuli maCvenebeli

statistikurad sarwmunod klebulobs (p<0,01).

aRniSnulidan gamomdinare, plaferoni lb-s samkurnalo efeqti

deqsametazonTan SedarebiT ufro Zlieradaa gamoxatuli. aRniSnuli

monacemebi imis sasargeblod metyvelebs, rom dakvirvebis bolo

vadisaTvis plaferoni lb RviZlis struqturis aRdgeniT procesebs

unda aCqarebdes.

bioqimiuri kvlevis Sedegebi RviZlis morfologiurma

gamokvlevamac daadastura. Con-A-s ineqciidan 8 sT-is Semdeg Tagvebis

RviZlSi mwvave anTebisaTvis damaxasiaTebeli morfologiuri

cvlilebebi aRiniSneba. hepatocitebis didi nawili vakuolur

distrofias ganicdis. amasTan erTad, fokaluri nekrozis kerebic

gvxvdeba. nekrozuli hepatocitebis citoplazma mkveTrad

eozinofiluria. RviZlis parenqimaSi limfoiduri rigis ujredebis

73

infiltratebi gvxvdeba. infiltracia gansakuTrebiT intensiuria

portul sivrceebSi.

deqsametazonis zemoqmedebis Sedegad eqsperimentis dawyebidan

merve saaTze hepatocitebis didi nawili vakuolur distrofias

ganicdis, Tumca wilakebis ZiriTadi trabekuluri struqtura

SenarCunebulia. portuli sivrceebis limfoiduri infiltracia

umniSvneloa. “slaj-fenomeni” sustad aris gamoxatuli. umetes

SemTxvevaSi eriTrocitebis konturebi Senaxulia. eqsperimentis am

vadaze Con-A-sTan erTad plaferoni lb-s Seyvanis SemTxvevaSic

hepatocitebis didi umravlesoba vakuolur distrofias ganicdis.

amasTanave, aRsaniSnavia, rom wilakis trabekuluri Seneba

SenarCunebulia. aseve iSviaTi da umniSvneloa limfoiduri

infiltraciis ubnebi.

Con-A-s Seyvanidan 24 sT-is Semdeg sakontrolo jgufis cxovelTa

RviZlSi cvlilebebi kvlav intensiuradaa gamoxatuli. hepatocitebis

umetesoba vakuolur distrofiasa da nekrozul cvlilebebs ganicdis.

deqsametazonis fonze mniSvnelovnad matulobs orbirTviani

hepatocitebis raodenoba. amave dros, aRiniSneba destruqciuli

cvlilebebis gaRrmaveba. plaferoni lb-s gavleniT hepatocitebis

umravlesoba kvlav distrofiul cvlilebebs ganicdis, magram

nekrozuli ubnebi ar aris.

Keqsperimentis dawyebidan 48 sT-is Semdeg sakontrolo jgufSi

zogierT areSi RviZlis normaluri trabekuluri struqtura vlindeba,

rac regeneraciuli procesebis gaaqtiurebaze miuTiTebs. amave vadaze

zogierTi normaluri hepatocitic Cndeba. deqsametazoniT mkurnalobis

SemTxvevaSi aRniSnuli suraTidan mniSvnelovani gansxvaveba ar

aRiniSneba. plaferonis gavleniT ki wilakis normuli Seneba ZiriTadad

aRdgenilia. nekrozuli ubnebi da limfoiduri infiltratebi saerTod

aRar SeiniSneba.

amgvarad, Con-A inducirebuli hepatitis dros plaferoni lb da

deqsametazoni RviZlis qsovils Rrma destruqciuli cvlilebebis

ganviTarebisagan icavs. aRsaniSnavia aseve, rom plaferoni lb

74

qsovilis struqturis aRdgenas aCqarebs, rac deqsametazonis

gamoyenebis SemTxvevaSi ar SeiniSneba..

rogorc ukve aRvniSneT, orive medikamenti - plaferoni lb da

deqsametazoni - RviZlis qsovils imunuri dazianebisagan icavs.

cnobilia, romHkortikosteroidebi aRniSnul efeqts citokinebis – il-1,

il-2, il-6, gama interferonisa da simsivnis nekrozis faqtori alfas -

genebis eqspresiis daTrgunvis gziT axorcieleben. savaraudoa, rom

plaferoni lb-s efeqtic mis mier citokinebis balansis SecvliT unda

iyos gamowveuli (Ciqovani, 1997), aqve unda aRiniSnos, rom

imunomamodulirebeli moqmedebis garda, plaferoni lb-s gamoxatuli

antioqsidaciuri da antiapoptozuri Tvisebebic aqvs (s. WoWua, 2006; m.

gongaZe, 2004). wina kvlevebSi naCvenebi iqna plaferoni lb–s Zlieri

antiapoptozuri Tviseba mwvave mZime formis hepatitis modelebSi da

klinikur gamocdebSi. kerZod, plaferoni lb ainhibirebs hepatocitebis

apoptozs LPS-iT inducirebul mZime hepatitis eqsperimentul modelSi.

metrevelis mier naCvenebi iqna, rom msubuqi da saSualo simZimis mqone

pacientebSi plaferoni lb-s gamoyeneba srul gamojanmrTelebas axdenda

96,9%-Si. ufro metic igi mniSvnelovnad amcirebda daavadebis

qronizaciis SesaZleblobas da xels uwyobda proteqtoruli

imunitetis Camoyalibebas (metreveli, 1999). am da sxva kvlevebis

safuZvelze Cvens mier SemoTavazebuli iqna hipoTeza, rom msubuqi

hepatitebis dros plaferonis damcvelobiTi Tviseba mis proapoptozur

unarTan aris dakavSirebuli.

cnobilia, rom kaspazebis gaaqtiureba apoptozuri procesis

damaxasiaTebeli niSania. apikaluri kaspazebis regulacia SeiZleba

gadamwyveti faqtori iyos am procesSi. kaspaza 8-s TviTgaaqtiureba

iwvevs kaspaza 3-s pirdapir gaaqtiurebasa da Bid–s gaxleCvas (Li et al.,

1998).

literaturuli monacemebis mixedviT, RviZlis imunopaTologiis

dros transkrifciuli meqanizmis funqciuri mdgomareoba gansazRvravs

imas, Tu romeli – kaspaza-3-s Tu alternatiuli – gziT warimarTeba

apoptozuri procesebi. Con-A–inducirebuli hepatitis dros kaspaza-3-s

75

aqtiurobis Sesaxeb monacemebi winaaRmdegobrivia. kerZod, erT-erTi

kvlevis mixedviT, kaspaza-3-s aqtiuroba mniSvnelovnad izrdeba (Ding et

al, 2004). xolo sxva gamokvlevis mixedviT, is ar vlindeba (Kunstle et al,

1999).

aqedan gamomdinare, kvlevis Semdeg etapze Cvens mier

Seswavlili iqna kaspaza-3-s aqtiuroba in vivo Con-A–s Seyvanis Semdeg

da masze imunomodulatorebis, kerZod, plaferoni lb–sa da

deqsametazonis gavlena. miRebuli Sedegebis Tanaxmad, eqsperimentis I

vadaze sakontrolo jgufis TagvebSi Con-A-s Seyvana kaspaza-3-s

aqtiurobis mkveTr matebas ganapirobebs da faqtiurad eqsperimentis

maqsimums aRwevs. amis Semdeg igi TandaTan klebulobs, Tumca 48 sT-

isTvis normas mainc ver ubrundeba (diag. 5).

plaferoni lb-s Seyvana eqsperimentis I da II vadaze kaspaza 3-s

aqtiurobaze ar moqmedebs da aRniSnuli maCvenebeli sarwmunod ar

gansxvavdeba sakontrolo jgufisgan. sakontrolo jgufTan SedarebiT

48 sT-ze kaspaza 3-s aqtiuroba mniSvnelovnad matulobs (p<0,01). maSin

roca, deqsametazonis gamoyeneba eqsperimentis I vadaze Trgunavs

kaspaza-3-is aqtiurobas sakontrolo jgufTan SedarebiT, xolo

eqsperimetis Semdeg vadebze aRniSnuli maCvenebeli sakontrolo

jgufis analogiuri maCveneblebisagan aRar gansxvavdeba.

kaspaza 3-s garda, Seswavlili iqna kaspaza 8-s aqtiurobac.

kaspaza 8-s aqtiurobis cvlilebebi kaspaza 3-s msgavsi aRmoCnda.

kerZod, Con-A–s Seyvanidan 8 saaTis Semdeg misi aqtiurobis mkveTri

mateba SeimCneva. 24 da 48 sT-isTvis ki kaspaza 8-s maCvenebeli

klebulobs, Tumca normas mainc ver ubrundeba (diag. 6).

plaferoni lb-s gamoyenebis SemTxvevaSi eqsperimentis I da II

vadaze kaspaza 8-s aqtiuroba kidev ufro izrdeba, Tumca igi

sarwmunod ar gansxvadeba sakontrolo jgufis maCveneblisgan. 48 sT-

ze ki sakvlev jgufTan SedarebiT kaspaza 8-is aqtiuroba

statistikurad sarwmunod matulobs (p<0,01). Cvens mier miRebuli

Sedegebis analizi cxadyofs, rom imunosupresanti deqsametazoni

kaspaza 8-s aqtiurobaze gavlenas ar axdens. deqsametazonis gavleniT

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kaspazebis inhibirebis tendencia vlindeba eqsperimentis pirvel

vadaze, Tumca is statistikurad sarwmunod ar gansxvavdeba

janmrTeli Tagvebis analogiuri maCveneblebisagan (diag. 6).

plaferoni lb ki proapoptozur Tvisebas avlens, rac msubuqi

hepatitis regeneraciis fazaSi kaspaza 3-sa da kaspaza 8-s aqtiurobis

mniSvnelovan matebaSi gamoixateba.

RviZlSi mimdinare apoptozuri procesebis Sesaswavlad

damatebiT gamoyenebul iqna TUNEL-is meTodi. miRebuli Sedegebis

safuZvelze dadginda, rom eqsperimentuli msubuqi hepatitis modelSi

plaferoni lb-s gamoyeneba 48 sT-ze sarwmunod zrdis apoptozuri

hepatocitebis raodenobas, Tumca igi ar moqmedebs eqsperimentis sxva

vadebze. aRsaniSnavia is faqti, rom imunomodulatori deqsametazoni

aseT efeqts ar avlens (cxrili 1).

uaxlesi monacemebiT Con-A-s mcire dozebis gamoyenebisas 48

saaTze xdeba bunebrivi T kilerebis raodenobis aRdgena, romelic

aucilebelia RviZlis normaluri regeneraciisTvis. SesaZlebelia

vivaraudoT, rom plaferoni lb xels uwyobs dazianebuli ujredebis

apoptozuri gziT mocilebas da rTavs iseT meqanizmebs, romlebic

imavdroulad hepatocitebis proliferacias aZliereben.

cnobilia, rom Warb apoptozs adgili aqvs mwvave da qronikuli

virusuli hepatitis, alkoholuri da araalkoholuri hepatitis,

RviZlis qolestaturi daavadebis, vilsonis daavadebisa da

“transplantanti maspinZlis winaaRmdeg daavadebis” (GVHD) dros. meore

mxriv, apoptozis deficiti asocirebulia RviZlSi da sanaRvle

sistemaSi simsivnis ganviTarebasa da progresirebasTan. ufro metic,

interferonis antivirusuli efeqti ganpirobebulia apoptozis

gamowveviT (Castelli et al, 1997).

Cveni kvlevis Sedegebi gvafiqrebinebs, rom plaferoni lb–s

gamoyeneba SesaZlebelia RviZlis sxvadasxva urTierTsawinaaRmdego

meqanizmiT mimdinare dazianebis dros. rogoricaa mwvave virusuli

hepatiti da simsivneebi. rogorc Cans, plaferoni lb apoptozuri

procesis modulirebas daavadebis formisa da simZimis mixedviT

77

axdens. RviZlis mZime dazianebis pirobebSi is antiapoptozur efeqts

avlens (s.WoWua da Tanaavt., 2006), xolo susti imunuri pasuxiT

ganpirobebuli RviZlis dazianebis SemTxvevaSi ki – proapoptozurs.

amdenad, SegviZlia davaskvnaT, rom plaferoni lb apoptozis

procesis modulatoria.

vinaidan imunuri sistemis funqcionireba ujredTaSorisi

komunikaciebiT ganisazRvreba, igi oqsidaciuri stresisadmi

gansakuTrebiT mgrZnobiarea. ujreduli komponentebis, upiratesad,

cilebis, lipidebisa da dnm-is endogenuri oqsidaciuri dazianeba

mravali qronikuli daavadebis paTogenezSi monawileobs. zogierTi

paTologiuri procesis, maT Soris, hepatitebis dros,DkavSiri

daqveiTebul antioqsidaciur statuss, oqsidaciur stressa da

avadmyofTa klinikur mdgomareobas Soris dadasturebulia. Tumca, ra

rols asrulebs oqsidaciuri stresi daavadebis ganviTarebasa da mis

progresSi, dRemde ucnobia.

cnobilia, rom oqsidaciuri stresis mimarT gansakuTrebiT

mgrZnobiarea apoptozis gamSvebi fermentebi kaspazebi. zogadad

miRebulia, rom mraval ujredSi reaqtiuli Jangbadis saxeobebis (ROS)

mcire koncentraciebi apoptozis iniciirebas iwvevs, maTi zeWarbi

raodenoba ki - nekrozul cvlilebebs ganapirobebs. kritikuli

cisteinebi sxvadasxva redoqs potencialiT xasiaTdebian, amitom

oqsidantebis mcire doza zogierT cilaSi Tiolis daJangvas iwvevs,

sxva cilebze ki ar moqmedebs.

amrigad, oqsidantebis mcire koncentracia apoptozis

ganviTarebas im redoqs statusis cvlilebebiT ganapirobebs, romlebic

xorcieldeba iseTi proapoptozuri faqtorebis gaaqtivebiT, rogoricaa

“c-Jun aminoterminaluri kinazis”. imavdroulad xdeba NF-kB-is

inhibireba, ise rom ar icvleba proapoptozuri kaspazebis redoqs

statusi. oqsidantebis maRali done ufro metad nekrozis ganviTarebas

ganapirobebs. isini ainhibireben kaspazebs, romlebic saWiroa apoptozis

gansaxorcieleblad, da ganapirobeben ujredebis nekrozul daRupvas

(Halliwell et al., 1990; Dypbukt et al., 1994). redoqs statusis Zlieri

78

cvlilebebi, rac ganpirobebulia wyalbadis peroqsidis raodenobis

matebiTYda glutaTionis SemcirebiT pirvelad kulturaSi

hepatocitebis nekrozs iwvevs. maSin, roca zomieri redoqs clilebebi

ar axdens gavlenas apoptozze. miuxedavad amisa, oqsidantebisa da

glutaTionis clilebebi aregulirebs TNF-isa da Fas-iT ganpirobebul

apoptozs. ufro metic, naCvenebia, rom xangrZlivi an Warbi oqsidaciuri

stresi kaspazebis aqtiurobis daTrgunvasa da Sesabamisad apoptozis

blokirebas axdens (Hampton et al., 1998).

imis gasarkvevad, Tu ra kavSiria msubuqi hepatitis dros

hepatocitebis apoptozsa da redoqs-potencialis cvlilebebs Soris,

amasTanave, imis dasadgenad, Tu ramdenadaa ganpirobebuli plaferoni

lb-s prokaspazuri aqtiuroba misi antioqsidaciuri unariT,

eqsperimentul cxovelTa sisxlSi SeviswavleT azotis oqsidisa da

glutaTion reduqtazas doneebis cvlileba, agreTve hepatocitebis

mitoqondriuli aqtiuroba.

sxvadasxva organoSi mimdinare metaboluri darRvevebi saboloo

jamSi sisxlSi ganviTarebuli cvlilebebiT aisaxeba, ris gamoc cocxal

organizmSi mimdinare metaboluri procesebis Sesaswavlad sisxli

yvelaze xelsayrel obieqts warmoadgens.

Cvens mier miRebuli monacemebis analizma cxadyo, rom Con-A-s

ineqciidan 8 saaTis Semdeg cxovelTa SratSi azotis oqsidis

Semcveloba statistikurad sarwmunod imatebs da eqsperimentis

bolomde amave doneze rCeba.

Con-A inducirebuli hepatitis dros deqsametazonis gamoyeneba

sisxlSi azotis oqsidis raodenobaze ar moqmedebs. aqedan

gamomdinare, igi antioqsidaciuri unaris SenarCunebas da lipidebis

zeJanguri Jangvis procesebis Sesustebas ver uzrunvelyofs.

aRsaniSnavia, rom plaferoni lb eqsperimentis II vadaze azotis

oqsidis koncentracias statistikurad sarwmunod Trgunavs (p<0,01).

eqsperimentis mesame vadaze ki plaferoni lb sapirispiro efeqts

avlens - sarwmunod zrdis azotis oqsidis raodenobas (diagrama 2).

eqsperimentis es vada RviZlis qsovilis regeneraciis fazas emTxveva.

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cnobilia, rom azotis oqsidi apoptozuri procesis

regulaciaSi monawileobs. igi avlens rogorc anti-, ise

proapoptozur moqmedebas. misi antiapoptozuri moqmedeba

xorcieldeba ori ZiriTadi meqanizmiT: ciklur

guanozinmonofosfatze damokidebuli da damoukidebuli. cikluri

guanozinmonofosfatis producirebis stimulirebiT NO iwvevs xsnadi

guanililciklazas aqtivacias, rac ujredebs Semdgomi apoptozisagan

icavs (Kim et al., 1997). es meqanizmi naklebadaa Seswavlili. meore,

ciklur guanozinmonofosfatze damoukidebuli meqanizmi, moicavs

lipidebis zeJangvis inhibirebas, peroqsil radikalis CaWeras, Bcl-2-is

gaxleCvisa da citoqrom C-s gamoTavisuflebis Semcirebas (Kim et al.,

1998) da iseTi cilebis induqcias, rogorebicaa Hsp70 da Bcl-2 (Mosser et

al., 1997; Genaro et al., 1995). garda amisa, NO damcvelobiT efeqts avlens

cilebis S-nitrozilirebiT. p50-isa da p65-is 62-e cisteinis S-

nitrozilirebiT NO-s ZaluZs NF-B-s aqtivaciis inhibireba. NO-s

donorebi ainhibireben TNF-s mier NF-B-s aqtivacias, rac IB--s

stabilizaciiTa da misi genis eqspresiis matebiT aris gamowveuli.

kaspazebi cisteinis maRali Semcvelobis gamo S-nitrozilirebisaTvis

idealur samizneebs warmoadgenen.

sainteresoa NO-s zemoqmedeba NF-B-s regulaciasa da anTebaze.

marTalia, misi moqmedebis meqanizmi bolomde ar aris garkveuli,

Tumca dadasturebulia misi rogorc pozitiuri, aseve negatiuri

gavlena aRniSnuli transkrifciuli faqtoris regulaciaze. es

moqmedeba damokidebulia azotis oqsidis koncentraciaze, redoqs

statusze, stimulsa da ujredis tipze (Kim et al., 1997).

endoTeliuri NOS–isa da/an neironuli NOS-is mier mcire

raodenobiT endogenurad warmoebul NO-s SeuZlia gadamwyveti roli

iTamaSos sxvadasxva stimulis dros NF-B-is aqtivaciis modulaciaSi.

in vitro da in vivo pirobebSi naCvenebi iqna, rom iNOS aferxebs TNF an

Fas-iT inducirebul apoptozs (Bogdan et al., 2001). ufro metic, in vivo iNOS

deficitur TagvebSi inhibirdeboda RviZlis regeneracia. garda amisa,

80

naCvenebi iqna (Sass et al., 2001), rom Con-A-inducirebuli RviZlis

dazianebis dros RviZlis qsovilSi ar warmoiqmneba 3-nitroTirozini,

azotis oqsidis mier qsovilis dazianebis markeri. sasma da kolegebma

(Sass et al., 2001) daadgines, rom hepatocitebSi inducibeluri azotis

oqsidis eqspresia ganapirobebs ujredebis nawilobriv dacvas.

Cvens mier miRebuli Sedegebis Tanaxmad plaferoni lb axdens

azotis oqsidis raodenobis modulirebas. kerZod, 24 saaTze preparati

amcirebs mis raodenobas (p<0,01) da amiT xels uSlis misi damangreveli

moqmedebis gavrcelebas. SedgomSi ki, 48 saaTisTvis igi axdens

inducibeluri azotis oqsidis raodenobis matebas zRvrul donemde

(p<0,01), riTac aCqarebs proliferaciul procesebs.

reaqtiuli Jangbadis saxeobebis ujredSida koncentracia mkacrad

kontroldeba dacvis mravalricxovani meqanizmebiT, maT Soris, ROS-is

skavenjeri fermentebisa da patara antioqsidanti molekulebis mier. am

antioqsidaciuri sistemebidan aRsaniSnavia glutaTioni da fermentebi,

romlebic oqsidaciuri dazianebisagan damcav faqtorebs warmoadgenen

rogorc cirkulaciaSi, ise sxvadasxva qsovilSi, maT Soris, RviZlSi

(Schwabe et al., 2006).

GSH–is saerTo ujredSida koncentracia mniSvnelovnad

cvalebadobs, gansakuTrebiT, RviZlSi. RviZlis GSH SeiZleba

dramatulad Semcirdes wamlis metabolizmis Sedegad, oqsidaciuri

stresis Semdeg an GSH–is sinTezSi Tandayolili defeqtebis gamo; GSH–

is dabali koncentraciebi gamovlenilia sefsisis, acetaminofenuri

intoqsikaciis, alkoholis qronikuli moxmarebisa da vilsonis mwvave

daavadebis dros (Uhlig et al., 1992). ufro metic, RviZlis GSH cirkadul

cvalebadobas ganicdis. RviZlis GSH–is raodenobis farmakologiuri

gazrda RviZls naklebmgrZnobiares xdis da icavs mravali pirdapiri

hepatotoqsinis zemoqmedebisgan (Prescott, 1982). kvlevebis mzardi

raodenoba miuTiTebs imaze, rom GSH–s ujredis dazianebaSi ormagi

roli gaaCnia.

81

naCvenebia, rom ujredis sikvdilis zogierT modelSi, sadac

apoptozi ZiriTad movlenas warmoadgens, GSH–is Semcireba damcav (e.i.

antiapoptozur) - da ara damamZimebel – efeqts qmnis. dReisaTvis

cnobilia, rom makrofagebis NO–inducirebuli apoptozi, T ujredebis

Fas–iT ganpirobebuli apoptozi da hepatocitebis citokin–gamowveuli

apoptozi in vivo damokidebulia Sesabamisi ujredebis ujredSida GSH–is

koncentraciebis sakmaris koncentraciaze (Hentze et al., 2000). wamoyenebuli

iyo hipoTeza, rom apoptozis ganmaxorcielebeli kaspazebis redoqs

mgrZnobiaroba pasuxismgebelia gamovlenil damcvelobiT efeqtze,

radgan GSH–is koncentraciebi Semcirebuli iyo. Tumca, GSH–is

gazrdil ujredSida koncentraciebs sxvadasxva ujredul xazSi

apoptozis gaZlierebac SeuZliaT (Uhlig et al., 1992). rac Seexeba in vivo

kvlevebs, GSH-is SemcirebiT gamowveuli damcvelobiTi efeqti naCvenebia

RviZlis apoptozuri kaspaz-damokidebuli dazianebis modelebSi (Hentze

et al., 1999).

kvlevis Semdeg etapze msubuqi hepatitis eqsperimentul

modelSi SeviswavleT glutaTion reduqtazas cvlileba cxovelTa

SratSi. rogorc miRebuli Sedegebis analizma cxadyo, Con-A-s

ineqciidan 8 sT-is Semdeg glutaTion reduqtazas aqtiuroba

statistikurad sarwmunod imatebs (p<0,01), 24 sT-sTvis igi mcirdeba,

Tumca sakontrolo maCvenebelTan SedarebiT isev momatebulia. 48sT-

sTvis glutaTion reduqtazas aqtiuroba normas ubrundeba.

plaferoni lb-s Seyvana statistikurad sarwmunod amcirebs

sakvlev jgufTan SedarebiT glutaTion reduqtazas raodenobas

(p<0,01). plaferoni lb mTeli eqsperimentis ganmavlobaSi glutaTion

reduqtazas koncentracias garkveul doneze inarCunebs, magram

normul maCveneblamde mis dayvanas ar axorcielebs (diagrama 3), rac

momatebuli proliferaciis periodSi saWiroa mitoqondriuli

funqciis Sesasruleblad.

plaferoni lb-s msgavsad, deqsametazonic glutaTion

reduqtazas maRali aqtiurobis SenarCunebas uzrunvelyofs.

82

aRsaniSnavia, rom Con-A-s ineqciidan 8 sT-is Semdeg plaferoni lb-s

gavleniT glutaTion reduqtazas aqtiuroba mniSvnelovnad mcirea

vidre deqsametazonis gamoyenebis SemTxvevaSi (p<0,01).

Cven mier gamovlenili monacemebi Seesabameba sxva kvlevis

Sedegebs, romlis mixedviTac, cirozsa da hepatocelularul

karcinomaSi sakontrolo qsovilTan SedarebiT glutaTion

reduqtazas aqtiuroba gazrdilia (Czeczot et al., 2006). ujredis doneze

GSH-is homeostazi uzrunvelyofilia biosinTezs, moxmarebas,

oqsidaciasa da eqsports Soris balansiT. homeostazis darRvevam ki

SeiZleba imoqmedos RviZlis unarze uzrunvelyos oqsidaciuri

dazianebisgan dacva (Fernandez-Checa et al., 1997).

kvlevis bolo etapze msubuqi hepatitis eqsperimentul modelSi

SeviswavleT hepatocitebis mitoqondriuli aqtiuroba MTT testis

saSualebiT. intaqturi Tagvebis hepatocitebis spontanuri

stimulaciis indeqsi (si) 1,14+0,2 udrida. Con A-Ti gamowveuli msubuqi

hepatitis dros 24 saaTisaTvis es maCvenebeli 1,5+0,3-is (sakontrolo

maCvenebelTan SedarebiT p<0,05), xolo 48 saaTisaTvis ki – 1,16+0,11-is

toli iyo.

plaferoni lb-s gavleniT Con A-inducirebidan 24 saaTis Semdeg

miRebuli sidide ar gansxvavdeba sakvlevi jgufis maCveneblisgan,

Tumca 48 saaTze is matulobs (2,06+0,12) da statistikurad sarwmunod

aRemateba intaqturi Tagvebis analogiur maCvenebels (p<0,01).

aRsaniSnavia is faqti, rom deqsametazonis SemTxvevaSi

mitoqondiriuli sunTqva daTrgunulia 48 saaTze (diagrama 4).

amrigad, miRebuli Sedegebi imis sasargeblod metyvelebs, rom

plaferoni lb mitoqondriul funqcias aZlierebs da hepatocitebis

proliferacias aCqarebs, riTac RviZlSi regeneraciuli procesebis

stimulirebas axdens.

83

daskvnebi

1. susti imunuri pasuxiT ganpirobebuli hepatitis dros

kaspazebiT gaSualebuli apoptozis intensivoba izrdeba.

2. Con-A inducirebuli msubuqi hepatitis dros plaferoni

lb-s gamoyeneba RviZlis qsovilis regeneraciis fazaSi

hepatocitebSi kaspazebis (3 da 8) aqtiurobas aZlierebs.

3. susti imunuri pasuxiT ganpirobebuli hepatitis

eqsperimentul modelSi imunomodulatori plaferoni lb

aZlierebs hepatocitebis apoptozs da Sesabamisad imunuri

pasuxis efeqts, riTac mcirdeba procesis qronizaciis

SesaZlebloba.


lb RviZlis qsovilis normaluri struqturis aRdgenas

aCqarebs.


lb sisxlSi azotis oqsidis Semcvelobas aregulirebs:

RviZlis maqsimaluri destruqciis fazaSi mas statistikurad

sarwmunod aqveiTebs, xolo regeneraciis fazaSi – zrdis.

6. RviZlis msubuqi imunuri dazianebis eqsperimentul

modelSi plaferoni lb oqsidaciuri stresis ganviTarebas

aferxebs.

7. plaferoni lb-s gavleniT hepatocitebis apoptozis

gaZliereba da amavdroulad maTi proliferaciuli

aqtiurobis mateba RviZlSi regeneraciuli procesebis

stimulacias ganapirobebs.

84

praqtikuli rekomendaciebi

1. susti imunuri pasuxiT mimdinare infeqciebis dros (B da C

hepatitis msubuqi formebi) samkurnalod rekomendebulia

plaferoni lb-s an proapoptozuri Tvisebis mqone sxva

imunomodulatoris gamoyeneba.

2. plaferoni lb-s moqmedebis meqanizmis ufro Rrmad

Sesaswavlad mizanSewonilia proanTebiTi da

anTebissawinaaRmdego citokinebis balansze misi zemoqmedebis

kvleva Con-A inducirebuli hepatitisa da autoimunur

daavadebaTa sxva eqsperimentul modelebSi.

3. plaferoni lb-Si Semavali imunomamodulirebeli

potencialiis mqone fraqciis identifikacia mizanSewonilia

Con-A inducirebuli hepatitis modelze.

85

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