B Y T TRNG I HC DC H NI ********** L DON TR NH GI TC DNG GII LO U THC NGHIM V MT S TC DNG HNG THN KINH KHC CA l - TETRAHYDROPALMATIN KHA LUN TT NGHIP DC S HNI- 2014 B Y T TRNG I HC DC H NI L DON TR NH GI TC DNG GII LO U THC NGHIM V MT S TC DNG HNG THN KINH KHC CA l - TETRAHYDROPALMATIN KHA LUN TT NGHIP DC S Ngi hng dn: 1.TS. Nguyn Hong Anh 2.Ths. Nguyn Thu Hng Ni thc hin: B mn Dc lc H NI- 2014 LI CM N Bng tt c s chn thnh v lng bit n su sc, em xin gi li cm n ti TS. Nguyn Hong Anh, Ths. Nguyn Thu Hng l nhng ngi thy, ngi c trctiphngdn,tntnhchbovgipemtrongsutqutrnhthc hin v hon thnh kha lun. EmxingilicmntiTS.NguynQucHuy,BmnThcvt,TS. NguynTrnLinh,BmnBoch,DS.PhmcVnh,BmnDclc, trngHDcHNicnhnggpvgipemtrongqutrnhthc hin ti. Em xin gi li cmn ti anhinh i , kthut vin ti bmn Dc lc trng H Dc H Ni nhit tnh gip v trc tip thc hin nhiu cng vic xuyn sut trong qu trnh tin hnh ti. Nhn y em cng xin gi li cm n ti: Cc thy c, cc anh ch k thut vin trong b mn Dc lc to mi iu kin, gip em c th hon thnh kha lun mt cch tt nht. CcthyctrongBangimhiunhtrng,ccphngban,ccbmn trng i hc Dc H Ni tn tnh dy d, gip em trong qu trnh hc tp ti trng.Emcngxinbytlngcmnchnthnhtinhngngithntronggia nh, nhng ngi bn lun kp thi ng vin, ng h em trong sut qu trnh hc tp ti trng cng nh trong thi gian thc hin kha lun. Em xin chn thnh cm n! H Ni, thng 5 nm 2014 Sinh vin L Don Tr DANH MC CC K HIU, CH VIT TT DANH MC CC BNG DANH MC CC HNH T VN ............................................................................................................ 1 CHNG 1: TNG QUAN ..................................................................................... 3 1.1. Lo u v ri lon lo u ...................................................................................... 3 1.1.1. Lo u ........................................................................................................... 3 1.1.2. Tnh hnh dch t ......................................................................................... 4 1.1.3. Phn loi ..................................................................................................... 4 1.1.4. Sinh ha thn kinh trong ri lon lo u ...................................................... 5 1.1.5. iu tr ri lon lo u bng cc thuc ha dc ........................................ 6 1.2. l tetrahydropalmain v tc dng dc l tm thn/thn kinh ........................ 8 1.2.1. Cu trc ha hc v tnh cht l ha .......................................................... 8 1.2.1.1. Cu trc ha hc .................................................................................. 8 1.2.1.2. Tnh cht l ha ................................................................................... 9 1.2.2. Ngun gc v chit xut ........................................................................... 10 1.2.3.Cc tc dng dc l hng thn kinh v tm thn ca tetrahydropalmatin. ............................................................................................. 12 CHNG 2: NGUYN LIU V PHNG PHP NGHIN CU .............. 16 2.1. Nguyn liu ..................................................................................................... 16 2.1.1. i tng nghin cu ............................................................................... 16 2.1.2. Ha cht, thit b ...................................................................................... 16 2.1.3. ng vt th nghim ................................................................................. 16 2.1.4 Chun b thuc th .................................................................................... 17 2.2. Phng php nghin cu ................................................................................ 17 2.2.1. S thit k nghin cu ......................................................................... 17 2.2.2. Ni dung nghin cu ................................................................................ 17 2.2.3. iu kin tin hnh th nghim ................................................................ 19 2.2.4. Liu lng s dng trong nghin cu ...................................................... 19 2.2.5. Cc m hnh dc l thc nghim s dng trong nghin cu ................. 20 2.2.5.1. nh gi tc dng gii lo u trn m hnh EPM ............................... 20 2.2.5.2. nh gi tc dng gii lo u trn m hnh ETM ............................... 21 2.2.5.3 nh gi nh hng trn vn ng t nhin trn m hnh OFT ........ 22 2.2.5.4. nh hng trn thi gian ng do thiopental...................................... 23 2.2.5.5. nh gi tc dng chng trm cm trn m hnh FST ..................... 23 2.3. X l s liu .................................................................................................... 24 CHNG 3: KT QU V BN LUN ............................................................ 26 3.1. nh gi tc dng gii lo u ca l tetrahydropalmatin ................................ 26 3.1.1 Trn m hnh ch thp nng cao, EPM ..................................................... 26 3.1.1.1. Thm d liu c tc dng gii lo u ca l - tetrahydropalmatin ....... 26 3.1.1.2. Tc dng gii lo u ca l tetrahydropalmatin s dng liu n v liu lp li 5 ngy............................................................................................ 27 3.1.2 Trn m hnh ch T nng cao, ETM ......................................................... 30 3.1.2.1. Sau khi s dng liu n l tetrahydropalmatin .............................. 30 3.1.2.2. Sau khi s dng liu lp li 21 ngy l tetrahydropalmatin ............ 31 3.2.nh hng trn hot ng t nhin v khm ph ca l-tetrahydropalmatin ... 32 3.2.1. Sau khi s dng liu n l tetrahydropalmatin ...................................... 33 3.2.2. Sau khi s dng liu lp li 5 ngy l tetrahydropalmatin ...................... 34 3.3. Tc dng an thn ca l tetrahydropalmatin ................................................. 34 3.4. Tc dng chng trm cm ca l tetrahydropalmatin ................................... 35 3.5. Bn lun .......................................................................................................... 36 3.5.1. V tc dng gii lo u ............................................................................... 37 3.5.2. V nh hng trn hot ng t nhin v kh nng khm ph ................ 39 3.5.3. V tc dng an thn .................................................................................. 41 3.5.4. V tc dng chng trm cm .................................................................... 41 KT LUN V XUT .................................................................................... 44 TI LIU THAM KHO DANH MC CC K HIU, CH VIT TT APAHip hi tm thn Hoa K (American Psychiatric Association) ASDRi lon stress cp (Acute stress disorder) BZDBenzodiazepine CLOClomipramin CREBProtein lin kt tng phn p ng vi AMP vng (Cyclic-AMP response element-binding protein) DZPDiazepam DSM-IV-TRDiagnostic and Statistical Manual of Mental Disorders, 4th Edition, Text Revision EPMM hnh ch thp nng cao (Elevated plus maze) ERKProtein chuyn gc phosphat iu ha tn hiu ngoi bo (Extracellular signal-regulated kinase) ETMM hnh ch T nng cao (Elevated T maze) FSTM hnh bi cng bc (Forced swim test) GABAGamma amino butyric acid GADRi lon lo u lan ta (Generalized anxiety disorder) ICD-10International Classification of Diseases, Tenth Revision IMAOMonoamino oxidase inhibitors IRTia hng ngoi (Infrared) OCDRi lon m nh cng ch (Obsessive compulsive disorder) OFTM hnh mi trng m (Open field test) PDRi lon hong s (panic disorder) PTSDRi lon stress sau chn thng (Post-traumatic stress disorder) RLLARi lon lo u SSRIc ch ti thu hi chn lc serotonin (Serotonin selective reuptake inhibitors) TCAsThuc chng trm cm 3 vng (Tricyclic antidepressants) THPTetrahydropalmatin WHOT chc Y t th gii (World health organization) DANH MC CC BNG BngTn bngTrang 1.1Kt qu nh lng THP (Nguyn Tin Vng)10 1.2Kt qu nh lng THP (L nh Mi)10 1.3Kt qu nh lng THP (Nguyn Quc Huy)11 3.1 Tc dng ca l tetrahydropalmatin v diazepam trn thi gian lu, s ln vo tay h v tng s ln lu li tay kn/tay h trn m hnh EPM 26 3.2 Tc dng ca l - tetrahydropalmatin v clomipramin s dng liu n trn cc ch tiu nh gi ca m hnh ETM 31 3.3 Tc dng ca l - tetrahydropalmatin v clomipramin s liu lp li 21 ngy trn cc ch tiu nh gi ca m hnh ETM 32 3.4 Tc dng ca l - tetrahydropalmatin v diazepam s dng liu n trn cc ch tiu nh gi ca m hnh OFT 33 3.5 Tc dng ca l - tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn cc ch tiu nh gi ca m hnh OFT 34 3.6 nh hng ca clomipramin, l tetrahydropalmatin thng qua cc ch tiu nh gi trn m hnh bi cng bc (FST) 36 DANH MC CC HNH HnhTn hnhTrang 1.1 Sc k ca dch chit alcaloid ton phn ca 6 loi thuc chi Stephania Lour. (UV 254 v 365 nm) 12 2.1M hnh ch thp nng cao20 2.2Dng c ch T nng cao21 2.3M hnh mi trng m22 2.4Dng c cho th nghim bi cng bc24 3.1 Tc dng ca l tetrahydropalmatin v diazepam s dng liu n trn thi lu li tay h 27 3.2 Tc dng ca l tetrahydropalmatin v diazepam s dng liu n trn s ln vo tay h v tng s ln lu li tay kn, tay h 28 3.3 Tc dng ca l tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn thi lu li tay h 29 3.4 Tc dng ca l tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn s ln vo tay h v tng s ln lu li tay kn, tay h 29 3.5 nh hng trn thi gian ng do thiopental ca diazepam (DZP) v l tetrahydropalmatin 35 1 T VN Rilonlou(anxietydisorders)lslosqumctrcmttnhhung xyra,ctnhchtvl,lplivkodigynhhngnsthchnghivi cuc sng ca ngi bnh. y l mt trong nhng ri lon tm l c tnh ph bin cao, c xuhng ngy cnggia tng, thng kt hp vi nhiurilon khc nh mt ng, trm cm, ri lon n ung, ri lon hnh vi [15] . Hin nay, phng php iutrrilonlouchyulphihpliuphptmlvithucgiilou (anxiolytics).Ccthucgiiloucngungchadcchnhcsdngl cc dn cht ca benzodiazepin, buspiron v cc thuc chng trm cm c ch chn lc ti thu hi serotonin hoc chng trm cm 3 vng [22]. Hn ch ln nht ca cc thuc c ngun gc ha dc l tc dng ph, kh nng l thuc thuc v phn ng caithucxyrakhidngiutr.Thmvo,githnhcaocaccthucny cng l vn cn quan tm khi s dng thuc hng ngy trong mt thi gian di cho bnh nhn. V vy, s dng cc thuc v sn phm c ngun gc dc liu hin ccoilhngtipcnbsungvthaythchoccthuccngungcha dc trong iu tr ri lon lo u [52]. l tetrahydropalmatin (l THP) l 1 alkaloid c tm thy nhiu loi thc vtkhcnhaunhngchyuctrongccloithucchiCorydalis(YanHuSuo) [46]vchiStephaniaLour.[21].TiVitNam,nm1941,BinhSangv Bonnet pht hin v chit xut c l THP t tn l rotundin, a Vit Nam thnh nc u tin chit xut c rotundin (1944: Nht, 1957: Cng ha dn ch c, 1960: Lin X, 1962: Trung Quc,). Theo Ng i Quang (1999), tc dng dc l ca l THP c nghin cu t thi Php thuc v c p dng iu tr t nm 1944 v trong sut cuc khng chin chng php i vi cc trng hp au tim, mt ng, hen, au bng; trong , tc dng r rt nht l tc dng an thn, gy ng. l THP c ngun gc t t nhin nn c nhng u im ni bt c tnh thp,sdungnpthuctt,mangligicngsinhl,khnggymtmi,nhc u nh cc thuc c ngun gc ha dc [14]. Gn y cc nh nghin cu quan 2 tm hn v cng c mt s nghin cu c cng b v nh gi v tm kim mc liu th hin tc dng gii lo u ca l THP [39] [62] [63]. gp phn nghin cu su hn v tc dng gii lo u v cc tc dng dc l tm thn kinh c lin quan ca l THP, to c s cho vic pht trin v ng dng thuc trong iu tr cc ri lon bnh l lin quan , ti c thc hin vi 2 mc tiu: 1. nh gi tc dng gii lo u thc nghim ca l tetrahydropalmatin 2. nh gi cc tc dng dc l hng thn kinh v tm thn c lin quan khc ti mc liu th hin tc dng gii lo u ca l tetrahydropalmatin. 3 CHNG 1: TNG QUAN 1.1. Lo u v ri lon lo u 1.1.1. Lo u Lo u (anxiety) l ri lon cm xc c trng bi cm gic lo s lan ta, kh chu, m h km theo cc triu chng thn kinh t ch nh au u, v m hi, hi hp,cmgicsitchtngc,khming,khchuvngthngvvbtrt, khngthngiynhocngynmtch.Loulmttnhiubong,bo trc mt nguy him sp xy ra v cho php con ngi s dng mi bin php ng u vi s e da [15]. Cn phn bit lo u bnh thng v lo u bnh l. S phn bit ny c th da trn kh nng kim sot lo u, thi gian ko di, cng v hnh vi km theo. i viloubnhthng,louthngxuthincngtnhhungvmtikhitnh hung c gii quyt; ng thi c th c iu tr hiu qu bng cch trn an hoc liu php tm l n gin. i vi lo u bnh l, lo u khng c nguyn nhn rrthocqumcsovimongi,cctriuchngthngnngvgynhiu kh chu, ko di dai dng, nh hng n hot ng hng ngy ca ngi bnh v c th km theo nhng ngh hoc hnh vi c v qu mc hoc v l (nh n trnh, hnhvicngch);ngthiviciutrcnphiciutrtheonguyn nhn sau khi c khm ton din v c nng v tm thn [15]. Ri lon lo u (RLLA, anxietydisorders)lmt trong nhng ri lon tm l c tnh ph bin cao, c xu hng ngy cng gia tng, thng kt hp vi nhiu ri lon khc nh mt ng, trm cm, ri lon n ung, ri lon hnh vi [15]. Trong , trm cm v lo u thng i km vi nhau;khong 3/4 bnh nhntrm cm c lo u km theo vtrn1/2 bnh nhn lo u cbiu hin trm cm rrt [23]. Nhiu bnhnhntmthnphnlitcthcmthylouddicngnhbnhnhn sng hoc sa st tm thn. Lo u cng c th va l nguyn nhn va l hu qu ca nghin ru hoc nghin ma ty. Nhiu bnh h thng cng kt hp vi lo u, trong trng hp ny lo u l do cc tc ng sinh l trc tip ca bnh h thng nhng 4 cng c th do iu tr hoc nhn thc tiu cc ca ngi bnh vbnh camnh [15]. 1.1.2. Tnh hnh dch t McdcSigmundFreudnghincutgnmtthktrcvitn gi lon thn kinh lo u (anxiety neurosis), cho n nay RLLA vn lun l mt lnh vcmim, thu hts quan tm ca nhiunhm nghin cu.Ch trong vng vi thp nin tr li y, cc nh khoa hc v thy thuc thc hnh xy dng cc tiu chun chn on, nh gi tl ca cc RLLA khc nhau, hiu c phn no c stml-sinhhccngnhphttrinmtsliuphphadcvtmlc hiu qu. Cc nghin dch t hc ti Hoa K cho thy c 4 ngi th c 1 ngi p ngcctiuchuncatnhtmtRLLAvtlbnhchungtrong1nml 17,7%,chiphchorilonlouchimkhong1/3chiphcanghnhtmthn, c bit lin quan nhiu n cc chi ph gin tip. Phi n thng b ri lon lo u nhiu hn phi nam vi t l bnh chung sut i ca hai phi ln lt l 30,5% v 19,2%, t l ny gim tng lp c thu nhp cao [23]. Ti Vit Nam, cha c iu tra dch t no c thc hin nh gi t l mc lo u v cc ri lon lin quan khc nhng vi tc th ha cao, nhp sng cng nghip v cng lm vic cng thng, s lng bnh nhn n khm vi ri lon lo u c xu hng ngy cng gia tng (ghi nhn ca bc s Nguyn Vn Nui, bnh vin Tm thn thnh ph H Ch Minh). 1.1.3. Phn loi Hin nay, da vo cc biu hin lm sng, cc nghin cu v dch t, di truyn v sinh hc cng nh s p ng vi cc phng php iu tr chuyn bit, ngi ta chia RLLA thnh nhiu loi khc nhau. S phn loi RLLA theo h thng phn loi bnh tt quc t ICD 10 ca T chc Y t th gii (WHO, 1992) v DSM-IV-TR caHipHitmthnHoaK(APA,2000)vcbngingnhau,trongri lon lo u c chia thnh cc loi nh sau [20] [23]: -Ri lon lo u ton th (generalized anxiety disorder, GAD) -Ri lon m nh cng ch (obsessive compulsive disorder, OCD) 5 -Ri lon hong s (panic disorder, PD) bao gm c hoc khng c m s khong rng (agoraphorbia) -Ri lon m s khong rng khng c tin s ri lon hong s -Ri lon m nh x hi (social phorbia) -Ri lon stress sau chn thng (post-traumatic stress disorder, PTSD) -Ri lon stress cp (acute stress disorder, ASD) 1.1.4. Sinh ha thn kinh trong ri lon lo u Cc nghin cu sinh l bnh v dc l ti thi im hin ti xut mi lin quan ca RLLA vi 3 c ch chnh [23]: -Kiunoradrenalin:lou,tngnhycmvphnngqumcvicctcnhn kchthchdnniuhaxung(downregulation)receptor2adrenergictrung ng. - Kiu benzodiazepin: triu chng lo u c th do gim hot ng h GABAA, cht dn truyn c ch ca thn kinh trung ng. - Kiu serotonin: cc triu chng ca ri lon hong s lin quann s thm ht serotonin dn ti gim hot ng ca con ng dn truyn serotonin. Trong ri lon lo u ton th (GAD), hiu qu iu tr ca cc benzodiazepin vbuspironhngccnghincusinhhcvocchthngdntruynthn kinh GABA v serotonin. Cc benzodiazepin (cht ch vn receptor benzodiazepin) c tc dng lm gim lo u trong khi flumazenil (cht i khng) li lm tng lo u. Mc d cha c bng chng v s bt thng ca cc receptor benzodiazepin trong rilonloutonth,mtsnghincutptrungvothychmlvngc mt receptor benzodiazepin cao nht trong no; cc vng no khc c cho l c lin quan n ri lon lo u ton th gm: cc hch y no, h vin v v no thy trn. Do buspiron l cht ch vn receptor 5-HT1A nn c gi thuyt rng c s btbnhthngtrongiuchnhhserotonergic.Cchthngdntruynthn kinhkhccngcnghincutrongrilonloutonthbaogm noradrenergic, glutamat v cholecystokinin [23]. 6 Trong ri lon hong s (PD), mt s nghin cu v c s sinh hc nhn thy cc triu chng ca ri lon ny c lin quan n cc bt thng sinh hc trong cu trcvchcnngno.Ccnghincunyangithuytchorngsri loniuchnh(dysregulation)cahthnkinhtrungngvngoibinclin quan n sinh l bnh ca ri lon hong s. H thn kinh t ch ca mt s bnh nhnmcchngrilonhongsccholbtngtrnglcgiaocm,thch ngchmvicckchthchmnhvpngqumcvicckchthchtrung bnh.Cchthnkinhchnhccholclinquanbaogmnoradrenergic, seroroninergicvacidgamma-aminobutyric(GABA).Srilonhotngh serotonergicrtrrttrongPD.Nhiunghincuviccthucchvni khng hn hp receptor serotonin lm tng lo u, cc png ny c th do s tngnhycmviserotoninsausynaptrongrilonhongs.Cngcbng chngtinlmsngchorngsgimchtdntruynloicchGABAccb tronghnhnhnybn,nogiavdiicthgyraccpngsinhl ginglou.Ccdliusinhhctptrungvonogia,hvin(cthlin quantilouvintng)vvnotrctrn(cthlinquantintrnhm nh).Trongsccchtdntruynthnkinhkhcnhauclinquan,h noradrenergiccchnhiunht,cbitvireceptor-adrenergictrc synap. Vai tr ca cc receptor ny c thm d qua th nghim dc l trn ng vt thc nghim vi clonidin (ch vn receptor 2) v yohimbin (i khng receptor 2) gy ra cc hot ng ging hong lon cc bnh nhn ri lon hong s [23]. 1.1.5. iu tr ri lon lo u bng cc thuc ha dc Hinnay,phngphpiutrrilonlouchyulphihpliuphp tm l vi thuc gii lo u (anxiolytics). Ba nhm thuc gii lo u c ngun gc ha dcchnhsdngtrongiutrbaogm:thucchngtrmcm,cc benzodiazepin v cc cht ch vn receptor 5-HT1A ca serotonin [4] [22] [23]. Cc thuc chng trm cm c ch chn lc ti thu hi serotonin (SSRI) -Thucsdng:fluoxetin,paroxetin,fluvoxamin,sertralin,citalopram, escitalopram. 7 -C ch: c ch chn lc ti thu hi serotonin v bc d tr tn cng t bo thn kinh trc synap, lm tng hot tnh serotoninergic trung ng. - Chnhiutrlou:CcSSRIhinlnhmthuclachnutaytrong nhiuthloubaogm:rilonmnhcngch(fluvoxamin,paroxetin, sertralin v fluoxetin), ri lon hong lon (paroxetin v sertralin), ri lon m nh x hi (paroxetin v sertralin), ri lon stresssau chnthng (paroxetin, sertralin v fluoxetin) do hot ph gii lo u rng v kh nng dung np tng i tt so vi cc nhm thuc khc. - Tc dng khng mong mun: ri lon tiu ha (bun nn, tiu chy, chn n, y bng),rilontnhdc(gimhammun,gimcckhoi),rilongicng,hi chngcaithucvtngtcthuc(cchenzymchuynhathucCYP2D6, CYP2C19, gyhi chng serotonin khidng cng tryptophan, lithi, IMAO v cc thuc chng trm cm 3 vng). Cc thuc chng trm cm ba vng- Thuc s dng: imipramin, clomipramin, doxepin. - C ch: c ch ti thu hi serotonin v noradrenalin v bc d tr tn cng t bo thn kinh trc synap, lm tng hot tnh serotoninergic v noradrenergic trung ng. - Chnhiutrlou:Ccthucchngtrmcm3vnghincxpvo nhm thuc la chn thay th (second line) trong iu tr ri lon lo u ton th, ri lon hong lon c m s khong rng v ri lon m nh cng ch. - Tc dng khng mong mun: gy ri lon thn kinh v tm thn (hoang tng, o gic, cmng, lo lng, l ln), khngcholinergic (gy h huyt p th ng, khming,tobn),rilonnitit(rilonkinhnguyt,gimtnhdc),c tnh cao hn SSRI trong trng hp qu liu, hi chng cai thuc khi ngng t ngt. Cc benzodiazepin (BZD) - Thuc s dng: diazepam, alprazolam, clonazepam. -Cch:lchtchvnreceptorbenzodiazepin,mtphncaphchpGABAA-knhCl-cnhiuith,hvinvvno.SgncaBZDvo receptor to thun li cho tc dng c ch ca GABA, tng s ln m knh Cl- qua trung gian GABA. 8 - Ch nh iu tr lo u: Cc BZD hin l nhm thuc la chn thay th trong mt s th lo u nh: ri lon hong lon (alprazolam v clonazepam), ri lon m s x hi(clonazepam).DotcdngbtlitrnnhmbnhnhncRLLAkmtheo trm cm hay nhm bnh nhn lm dng/nghin ru nn v tr ca BZD trong iu trbhnchhnsoviccSSRI.KhcviSSRIvthucchngtrmcm3 vng, BZD khi pht tc dng rt nhanh, do thng c ch nh phi hp iu tr vi SSRI trong giai on u.- Tcdngkhngmongmun:anthn,tnthngvnhnthc(khtptrung, gimtrnh,qunthngtinmi),mtphihpvnng,suyhhp,lmdng thuc (ti pht, bt li v hi chng cai thuc). Cht ch vn tng phn receptor 5-HT1A (buspiron)- Ch nh iu tr lo u: Ch c hiu qu trong iu tr ri lon lo u ton th. Do khi pht tc dng chm ng thi khng chc chn v hiu qu chng ti pht khi dng lu di nn buspiron c coi l thuc la chn thay th. - Tc dng khng mong mun: bun nn, chng mt, au u, mt mi, bun ng (lc bt u dng thuc), i khi gy ri lon vn ng, chng my c. Nhn chung, buspiron dung np tt hn so vi BZD, t gy bt li hn trn i tng bnh nhn cao tui. 1.2. l tetrahydropalmain v tc dng dc l tm thn/thn kinh 1.2.1. Cu trc ha hc v tnh cht l ha 1.2.1.1. Cu trc ha hc TiVitNam,nm1941,BonnetvBinhSangchittcbnhvi StephaniarotundaLour.mtalkaloidttnlrotundin.Nm1965,alkaloidny c xc nh l l-tetrahydropalmatin [17]. Cng thc cu to [6]. 9 -Tnkhoahc:5,8,13,13atetrahydro2,3,9,10tetramethoxy6H dibenzo[a,g] quinolizin. -Tn khc: rotundin, gindarin, caseanin, hyndarin. -Cng thc phn t: C21H25NO4 1.2.1.2. Tnh cht l ha Tnh cht vt l [6] -ltetrahydropalmatinlmtalkaloidthrn,cmutrng,khngmi, khng v. B chuyn thnh mu vng khi tip xc vi nh sng hoc tip xc vi nhit. -V tnh tan, l tetrahydropalmatin khng tan trong nc, t tan trong ethanol v ether, tan trong chloroform, d tan trong acid sulfuric long. Khi tn ti dng mui, l tetrahydropalmatin d tan trong nc. -Nng sut quay cc: []D20 = - 2910 (C = 0,8 ; ethanol 950). -im chy: mp = 1410C 1440C. -Ph hng ngoi (IR) c trng. Tnh cht ha hc [5] -ltetrahydropalmatinmangctrnghatnhcaNbc3:phnngvi thuc th chung ca alkaloid, to mui vi cc acid v c. -Mch kp: oxy ha l tetrahydropalmatin (mu trng) bng iod s to thnh palmatin c mu vng.

l tetrahydropalmatin (mu trng)palmatin ( mu vng) Boqun:ltetrahydropalmatincboquntronglthytinhknti mu, ni kh ro, trnh nh sng. 10 1.2.2. Ngun gc v chit xut Ngun gc ltetrahydropalmatinl1alkaloidctmthynhiuloithcvtkhc nhau nhng ch yu c tm thy trong cc loi thuc chi Corydalis (Yan Hu Suo) [54] [46] v chi Stephania Lour. [21]. l tetrahydropalmatin c phn lp t 16 loithucchiStephaniatrnthgiiv7loiVitNam,vlhotchtchnh cphnlptnhiuloinhttrongnhmalkaloidkhungprotoberberin,cng nhlthnhphnhahcchnhcachiStephaniaLour.[11].TiVitNam,l tetrahydropalmatin ch yu c chit xut t cc loi thuc chi Stephania Lour. Nm2000,NguynTinVngxcnhhmlngtetrahydropalmatin (THP) t 3 loi bnh vi nghin cu [18]: Bng 1.1. Kt qu nh lng THP (Nguyn Tin Vng) STTLoiHm lng THP (%) 1S. glabra (Roxb.) Miers0,59 2S. kuinanensis H.S.Lo3,06 3Stephania sp3.1,52 Nm2005,LnhMikhosthmlngTHPtrong5loibnhvi nghin cu [13]: Bng 1.2. Kt qu nh lng THP (L nh Mi) STTLoi Hm lng THP (%) Hm lng THP/alcaloid ton phn (%) 1S. brachyandra Diels.5,150,1 2 S. glabra (Roxb.) Miers 2,150,0 3 S.sinica Diels. 1,446,4 4 S.kwangsiensis H.S.Lo 1,850,5 5 S.hainanensis H.S.Lo 0,618,3 11 VXunGiangxcnhhmlngTHPvpalmatintrongloiS. viridiflavens H.S. Lothu hi Sn La l: 0,63% v0,32% [10] . Phm Th Duyn (2004) nghin cu nh lng THP bng phng php sc k lng hiu nngcao (HPLC) cho thy hm lng THP trong loi S. dielsiana (thu hi Ba V-H Ty), loi S. viridaflavens (thu hi Sn La) v Stephania sp. (trong vin Balok) ln lt l: 0,40%; 0,62% v 1,63% [7] [8]. Bi Th Bng xc nh hm lng THP trong cc loi S. brachyandra v S. sinica ln lt l: 3,55% v 1,31% [2]. Nm 2010, Nguyn Quc Huy tin hnh nh lng THP trong 3 loi nghin cu [11]. Bng 1.3. Kt qu nh lng THP (Nguyn Quc Huy) STTLoiHm lng THP (%) 1S. brachyandra Diels.3,69 2S. dielsiana Y. C. Wu.0,40 3S. sinica Diels.2,43 C th thy r c s khc bit rt ln v hm lng alcaloid v THP gia cc loi chi Stephania Lour. pht hin ti Vit Nam. Kt qu kho st bc u trn sc k lp mng ca Quyn v Nguyn Quc Huy cng th hin r iu ny: 12 Hnh1.1. Sc k ca dch chit alcaloid ton phn ca 6 loi thuc chi Stephania Lour. (UV 254 v 365 nm) BR: S. brachyandra, GL: S. glabra, KW: S. kwangsiensis, SI: S. sinica; SP1: Stephania sp.1 (Mu Ha Bnh), DI: S. dielsiana, R: L-tetrahydropalmatin Dung mi trin khai sc k: Cloroform : Aceton : NH4OH [8 : 2 : 0,1] 1.2.3.Cc tc dng dc l hng thn kinh v tm thn ca tetrahydropalmatin. mcphnt,tetrahydropalmatin(THP),mtalcaloidctmthy huhtccloithucchiStephania,cilcgncaovireceptor1Av2A-adrenergic, receptor D1, D2 v D3 ca dopamin, receptor5-HT1A, 5-HT1D, 5-HT4 v 5-HT7 caserotonin[42][43].Qua,lmcnkitcccpdntruynthn kinhcanoradrenalin,dopaminvserotonintronghthngthnkinhtrungng [43]. Tc dng an thn ca THP c cho rng c th do hot ng i khng ca ntrnreceptorD1,D2vD3[38].TcdngikhngcaTHPtrnreceptor dopamin gim dn theo th t D1, D2, D3. Tc dng gim au ca THP c chng minhcngclinquantitcdngcchreceptordopaminD2[26].THP c chng minh rng c tc dng h huyt p ng mch v gim nhp tim thng qua qu trnh gii phng ca h thn kinh serotonergic vng di i [27]. c nghincuchngminhskhcnhautrongcchtcdngca2ngphni nhaucadltetrahydropalmatinl:d-tetrahydropalmatin(dTHP)vl 13 tetrahydropalmatin (l THP), trong khi d THP tc dng nh mt cht lm cn kit nngdopaminthlTHPlilchtikhngvidopamintrnreceptorca chttrunggianhahcny[61].THPctcdngikhngreceptor1-adrenergic, ch vn trn receptor 2-adrenergic v l cht iu ha d lp th dng tnh ca receptor GABAA [36] [59]. THP i khng tc dng hot ha cc receptor serotonin,noradrenalinvdopamincacocain.Tcdngtrnreceptor5-HT1Av -adrenergicgipgimnhtcdngngoithpdoikhngdopamin[47].THP ctcdnggimcctriuchngtrongqutrnhcainghincocainvherointrn thcnghimchutcng[47][64],lmgimsthmmunvtnhtrngti nghintrongqutrnhcainghinheroin[60].THPcchsphthuctmthn gyrabioxycodonthngquanhhngtisphosphorylhaCREB(cyclic-AMPresponseelement-bindingprotein)vERK(Extracellularsignal-regulated kinases) nhn vng v hi hi m (hippocampus) chut cng [44]. Tc dngc ch thnkinh trung ng ca dl-tetrahydropalmatin (THP),mt alkaloid c phn lp t nhiu loi trong chiStephania c chng minh trn cc m hnh dc l thc nghim: NguynTinVngtinhnhthmdtcdngcal-THPtrninno th. Tt c cc th c tin hnh t in cc vo vng cm gic, vn ng v th lithnno.Saukhichothungl-THPviliu100mg/kgthtrngtrong7 ngy, thnh phn cc sng in no trongv no vng cm gic-vn ng v trong th li thn v no c nhng bin ng r. S tng thnh phn sng chm delta v gim thnh phn sng nhanh beta chng t rng l-THP c tc dng tng cng c ch t bo thn kinh v bn cu i no v th li thn no [18]. V tc dng gii lo u, nghin cu ca Leung v cng s cho thy liu thp (0,5 10 mg/kg), THP c tc dng gii lo u r rt, tc dng an thn v gin c ch thhinliucao(50mg/kg).Tcdnggiiloucatetrahydropalmatinbi khngbiflumazenil,mtchtcnhtranhvtrgncabenzodiazepintrn GABAA receptor, chng t c ch gii lo u ca THP thng qua, t nht mt phn, tngtcvivtrgncabenzodiazepintrnreceptorGABAA [36][39].Cng 14 trong nghin cu v THP, Hong Xue v cng s pht hin: dl THP th hin tc dng gii lo u r rt cc mc liu 0,1 mg/kg n 50 mg/kg tnh theo trng lng c th chut [63], trong khi l THP cc mc liu t0,1 mg/kg n 2,5 mg/kg tnhtheotrnglngchutthhintcdnggiilourrt.Tcdnganthn xut hin mc liu cao (5 mg/kg) [62], ng thi c ch tc dng c th lin quan nkhnngdlTHPvl-THPlnhngchtchvntivtrgn benzodiazepin trn receptor GABAA c xut [62] [63]. Vtcdnganthn,trnmhnhtngcngthigianngdothiopental, PhmDuyMaivcngschothygindarinhydrochlorid(l tetrahydropalmatin hydrochlorid) mc liu 30 mg/kg th trng chut ko di thi gianngdothiopentalln1,5lnsovilchng[12].CngtheoNguynTin Vng,lTHPmcliu40mg/kgthtrngchutlmtngthigianngdo thiopentalln8lnsovinhmchng[18].TheoPhmVnVngvcngs, nghin cu tc dng gy ng ca siro laroxen (rotundin sulfat) cho thy mc liu 120mg/kg th trng chut c tc dnggyng v kodi thi gian ng tng t diazepam [19]. Vnhhngtrnhotngtnhin,KaiYuevcngschothyl THPmcliu5mg/kgtnhtheothtrngchutlmgimrrtkhnnghot ngtrnchutcng,trongkhiccmcliuthphnkhngchothynhhng ny[64].NghincucaHongXuevcngschothymcliulmgimhot ng t nhin ca dl THP v l THP tng ng l: t 0,1 mg/kg n 100 mg/kg v t 5 mg/kg n 40 mg/kg tnh theo trng lng c th chut [62] [63]. Kt qu nghin cu ca Phm Vn Vng v cng s cng cho thy Laroxen (rotundinsulfat)mcliu20mg/kgchothytcdngcchhunghncachut vi t l phn trm c ch hung hn l 36,91% tng t diazepam. mc liu 60 mg/kg,rotundinsulfatcngthhintcdngchngcogitgyrabicorazolv th hin tc dng gim au mc liu 30 mg/kg [19]. ng dng trong iu tr, l tetrahydropalmatin t lu c s dng iu tr mt s ri lon bnh l tm thn v thn kinh. Ti Trung Quc, hiu qu ca l 15 THP nh mt thuc gim au khng opioid, mt hot cht an thn/gii lo u, h tr iu tr cai nghin dn n s chp thun trong ch nh iu tr cc bnh l tng ng [59]. Ti Vit Nam, nhiu sn phm cha THP lu hnh trn th trng vi cc bit dc: Rotundin, Rotundin 30, Rotunda... c s dng vi ch nh iu tr an thn gy ng, gim au, h tr iu tr cai nghin. 16 CHNG 2: NGUYN LIU V PHNG PHP NGHIN CU 2.1. Nguyn liu 2.1.1. i tng nghin cu l tetrahydropalmatin (Rotundin) t tiu chun kim nghim vi hm lng 98,4%,ccungcpbicngtyCPhnDcphmTmHiuc.Mul- tetrahydropalmatinckimnghimtiphngkimtrachtlng,cngtyc phn Traphaco theo tiu chun c s CP 1998. 2.1.2. Ha cht, thit b Diazepam(bitdcSeduxencaGedeonRichter,Hungari),clomipramin (bitdcClomidep25mgcaCngtySunPharmaceuticalIndustries,n), thiopental natri ca Rotexmedica (c) do cng ty c phn Dc phm TW2 phn phi. Dung dch Tween 80 1%. My nghin bi siu mn Retsch mm20 sn xut bi hng Retsch GmbH & Co RG ca c, b mn Bo Ch, trng i hc Dc H Ni. Bm, kim tim nha, ng h bm giy, nhit k, bng n 60W, n bn, cn phn tch, cn k thut. Cccm100,250ml,bnhnhmc10,20,25,50v100ml,chy,ci, micropipet 0 5000 l v mt s dng c dng trong chn nui v pha ch ca b mn Dc lc, trng i hc Dc H Ni. 2.1.3. ng vt th nghim +Chutnhttrnggingc,chngSwissalbinokhemnh,khilng trung bnh 18-22 g do Vin V sinh Dch t Trung ng cung cp.+ Chut cng trng c ging ci, chng Wistar, khi lng 200-250 g do Hc vin Qun Y cung cp. Chut c chia l ngu nhin 8-10 con/l, nui trong iu kin nhit 241oC, nh sng t nhin, c nui bng thc n chun, ung nc t do. 17 2.1.4 Chun b thuc th Cc thuc th c s dng di dng hn dch: diazepam, clomipramin, l tetrahydropalmatin c phn tn ng nht trong Tween 80 1% t c nng mong mun. Tt c cc thuc u c s dng theo ng ung. 2.2. Phng php nghin cu 2.2.1. S thit k nghin cu 2.2.2. Ni dung nghin cu Nghin cu c thit k thnh 3 t th nghim vi cc ni dung sau y: t 1: Th nghimtrn chut nht,thm d liu c tc dng gii lo u v an thntrnccmhnhEPMvmhnhtngcngthigianngdothiopental tng ng. Chn ra cc mc liu c tc dng tin hnh cc th nghim tip theo, th nghim c thc hin theo s sau: l - tetrahydropalmatin Tc dng gii lo u Tc dng gii lo u, m hnh ch thp nng cao (EPM) Tc dng gii lo u ton th v chng hong s, m hnh ch T nng cao (ETM) nh hng trn hot ng t nhin ca chut, m hnh mi trng m (OFT) Tc dng an thn, tng cng thi gian ng do thiopental Tc dng chng trm cm, m hnh bi cng bc (FST) 18 3-5 ngyBt u 5 ngy Kt thc thch nghi iu kincho ung thuc lin tc th nghim Ung liu u tin 5 ngy ung thuc M hnh EPM Tim tnh mch thiopental t 2: Th nghim trn chut nht,nh gi tc dnggii lo u s dng liu n v s dng liu lp li l tetrahydropalmatin trn m hnh EPM; nh gi nh hngtrnhotngtnhincachutsdngnliuvliulplical tetrahydropalmatin trn m hnh OFT, th nghim c thc hin theo s sau: 3-5 ngyBt u 5 ngy Kt thc thch nghi iu kincho ung thuc lin tc th nghim Ung liu u tin 5 ngy ung thuc M hnh EPMM hnh EPM M hnh OFT M hnh OFT t 3: Th nghim trn chut cng, nh gi tc dng gii lo u ton th v tc dngchnghongscal-tetrahydropalmatintrnmhnhETM;nhgitc dngchngtrmcmcaltetrahydropalmatintrnmhnhFST,thnghim c thc hin theo s sau: 19 5- 7 ngyBt u 21 ngy 4 ngy Kt thc thch nghicho ung thuc lin tctip tc ung thuc iu kin th nghim Ung liu u tin 21 ngy ung thuc25 ngy ung thuc M hnh ETM M hnh ETM M hnh FST 2.2.3. iu kin tin hnh th nghim Tt c cc th nghim u c tin hnh trong phng ring bit,yn tnh ti khu chn nui ng vt th nghim ca trng i hc Dc H Ni. Thi gian tin hnhthnghimtrongkhongt8h-17h.VimhnhEPMvmhnhOFT, cng chiu sng trong thi gian lm thc nghim c duy tr bng 1 bng n 60W.VimhnhETM,cngchiusngtrongthigianlmthcnghim c duy tr bng 2 bng n 60W. Vi m hnh FST, cng nh sng duy tr l nh sng t nhin. 2.2.4. Liu lng s dng trong nghin cu Trongthnghimthmdliuctcdnggiilouvanthn(t1),s dng7mcliucal-tetrahydropalmatinl0,03mg/kg,0,1mg/kg,0,3mg/kg, 1mg/kg, 3 mg/kg, 10 mg/kg v 100 mg/kg tnh theo trong lng c th chut. Cc thnghimsau(t2vt3)sdngmcliuchiuqucxcnh trong t 1. Liu ca cht i chiu diazepam c chn mc 2 mg/kg th trng chut theo Dng Phc An v cng s [1], mc liu ny ph hp vi mc liu ca diazepam dng iu tr lo u trn ngi trng thnh dng ng ung (4- 15 mg/ngy)[3],quyiraliucachutnhttrng.Mcliucachngdng clomipramincsdngtrongnghincul25mg/kg,sdngdingy(21 ngy) theo Poltronieri v cng s [50]. Mc liu ny ph hp theo mc liu quy i t mc liu dng trn ngi iu tr ri lon hong s sang liu trn chut cng 20 [3].CcthucthcphntntrongTween801%tohndchchochut ung. 2.2.5. Cc m hnh dc l thc nghim s dng trong nghin cu 2.2.5.1. nh gi tc dng gii lo u trn m hnh EPM M hnh ch thp nng cao theo m t ca Lister [41]. Hnh 2.1. M hnh ch thp nng cao Dng c th nghim Dng c bng g gm 2 tay kn kch thc 30 x 5 x 15cm (di x rng x cao), 2 tay h kch thc 30 x 5 x 0,2 cm, 2 tay kn v m t vung gc vi nhauv t cchnnnh60cmtothnhhnhchthp,cngchiusngtrongthigian lm thc nghim c duy tr bng mt bng n 60W. Phng php tin hnh Chutcchiangunhinthnhcclvchoungthucthmingy1 ln vo 8h. Trongccngylmthnghimtrnmhnh,saukhiungthuc60pht, chutctnhnhngvovngtrungtm,mthngvcnhtayh,sau c t do di chuyn khm ph trong 5 pht. Chut c coi l trong 1 cnh tay khic4chnunmtrongcnhtay.Saukhitinhnhthnghimvimi chut, dng c th nghim c lau sch bng khn tm cn trc khi tin hnh vi 21 chut tip theo trnh lu gi mi. Ch tiu nh gi: ghi nhn thi gian lu li tay h, s ln ra vo cc vng cnh tay (ng v/hoc h) ca chut. 2.2.5.2. nh gi tc dng gii lo u trn m hnh ETM M hnh ch T nng cao theo m t ca Graeff v cng s [35]. Dng c th nghim Hnh 2.2. Dng c ch T nng cao Dng c th nghim bng g c cu to l hnh ch T gm 1 cnh tay kn kch thc 50 x 12 x 40cm (di x rng x cao) v 2 tay h kch thc 50 x 12 x 1 cm, tay kn c t vung gc vi 2 tay h. B dng c c t cch nn nh 50cm, cng chiu sng trong thi gian lm thc nghim c duy tr bng 2 bng n 60W. Phng php tin hnh Chutcchiangunhinthnhcclvchoungthucthmingy1 ln vo 8h. Pre-test: trc ngy th nghim 1 ngy, chut c phi nhim vi mt trong hai cnh tay h trong vng 30 pht, mt thanh g chn ngang trc li vo ca cnh tay kn v mt thanh chn ngang u gn cnh tay h m bo chut c tch bitviphncnlicadngc.Chutcungthucsaukhiphinhim30 22 pht (khng cho chut ung thuc sau pre-test i vi th nghim nh gi tc dng sau khi ung liu u tin). T-test:ccthnghimctinhnh24hsaupre-test.Trongngylmth nghim, sau khi cho chut ung thuc 1h, chut c t vo trong mt chic lng schvkhchunbtrctrong2pht.Sauchutctvocuicnh tay kn, mt hng v trung tm t do khm ph m hnh. Ghi nhn thi gian k tkhitchuttikhichutritayknbngc4chn(thigiantimtngnn, baselinelatency),saughinhnlithigiannytrong2lntchutvom hnhlintip(ntrnh1vntrnh2,avoidance1vavoidance2).Sauhun luyn n trnh, chut c t li vo m hnh vo phn cui tay h bn phi, mt hngvphatrungtmvghinhnthigianchutritaynybngc4chn (chytrn,escape).Thigiantiachomithnghiml5pht.Giaccth nghim, dng c c lau sch bng cn trnh lu gi mi. Trong khong thi gian 30 giy gia cc th nghim chut c t li vo lng chun b sn. 2.2.5.3 nh gi nh hng trn vn ng t nhin trn m hnh OFT Xc nh theo m t ca Todd v cng s [56]. Dng c th nghim Hnh 2.3. M hnh mi trng m 23 Dngcthnghimhnhhpvungbngg,bntrongcptmhpkim nhmnha(aluminium)muxmkchthc60x60x40cm(dixrngxcao) bntrongsnphnthnh16vungnhbngnhau(4x4)vtngngvi2 vnglvngtrungtm(4nmgia)vvngvinngoi(12ngoicnli), cng nh sng lm th nghim duy tr bng bng n 60W. Phng php tin hnh Trong ngy lm th nghim, sau khi chochut ungthuc1h, nhnhng t chut vo chnh gia vng trung tm ca m hnh u trng m hng v pha 1 cnh ca m hnh; quay video ghi nhn cc hot ng ca chut trong vng 5 pht. Sau khi tin hnh th nghim vi mi con chut, dng c th nghim c lau sch bng khn tm cn trc khi tin hnh vi chut tip theo trnh lu gi mi. Ch tiu nh gi: S ng k i qua, s ln ng ln bng 2 chn sau ca chut, s ln vo trung tm, thi gian trung tm. Trong : chut c coi l i qua 1 ng k hay vo vng trung tm khi c 4 chn ca chut i qua ng k v vo vng trung tm; chut ctnh l1 ln ng lnbng 2 chn sau khi chut chuyn t trng thi ng bng 4 chn sang ng bng 2 chn sau (2 chn trc c th chm thnh hoc l khng). 2.2.5.4. nh hng trn thi gian ng do thiopental Xc nh theo m t ca Trung m v Th Phng [9]. Chutctimtnhmchuithiopentalliu40mg/kg.tchutnm nghing, thi gian ng do thiopental c tnh t khi chut mt phn x thng bng cho n khi chut c phn x thng bng tr li. 2.2.5.5. nh gi tc dng chng trm cm trn m hnh FST Xc nh theo m t ca Slattery v Cryan [53].Dng c th nghim Dng c l mt bnh thy tinh trong sut hnh tr c chiu cao 60 cm, ng knh 25 cm, chiu cao mc nc 30 cm, nhit nc c duy tr trong khong 23 25oC. 24 Hnh 2.4. Dng c cho th nghim bi cng bc. Phng php tin hnh Pre-test:trcngythnghim1ngy,chutcphinhimvimhnh bng cch cho chut bi trong m hnh 15 pht. Sau , cho chut ra ngoi, lau kh bng khn sch v si m bng n si t 60W. Sau khi chut phi nhim vi m hnh 30 pht cho chut ung thuc. Thnghim:Thnghimctinhnhsau24htkhithchinpre-test. Trong ngy th nghim, sau khi cho chut ung thuc 60 pht, th chut nh nhng vomhnh.Thigianquanstchomithnghimtrongvng5pht.Sauth nghim vi mi chut, trng sch dng c v thay nc mi trnh lu tr mi. Ch tiu quan st: thi gian cho mi trng thi tro hoc ln, bi v bt ng. 2.3. X l s liu D liu c lu tr, phn tch bng phn mm SPSS 16.0. Kt qu c biu din di dng gi tr trung bnh sai s chun (X SE). QuaphntchkimchunKolmogorow-Smirnowchothymtsdliukhng 25 tun theo phn b chun. V vy, mu khng tun theo phn b chun v cc mu m bin c tnh cht khng lin tc c kim nh bng test phi tham s Kruskal Wallis sau l Mann Whitney U test so snh s khc bit gia cc l. Vi cc mutuntheophnbchun,sdngkimnhOnewayAnovavihukim (post hoc) Dunnette T3 so snh s khc bit gia cc l. S khc bit c coi c ngha thng k khi p < 0,05. 26 CHNG 3: KT QU V BN LUN 3.1. nh gi tc dng gii lo u ca l tetrahydropalmatin 3.1.1 Trn m hnh ch thp nng cao, EPM 3.1.1.1. Thm d liu c tc dng gii lo u ca l - tetrahydropalmatin Thnghimthchintrn7mcliucaltetrahydropalmatin(t0,03 mg/kgn100mg/kg)vimcchthmdliuctcdnggiilou.Ktqu thuctrnthigianlutitayh,slnvotayhvtngslnlulitay kn/tay h ca cc l chut th nghim trong m hnh ch thp nng cao c trnh by trong bng 3.1. Bng 3.1. Tc dng ca l tetrahydropalmatin v diazepam trn thi gian lu, s ln vo tay h v tng s ln lu li tay kn/tay h trn m hnh EPM Ln Thi gian lu ti tay h (giy) S ln vo tay h Tng s ln lu li tay kn/tay h NaCl102,20 1,140,50 0,319,10 1,85 DZP 2 mg/kg 10 46,50 15,11 p = 0,019 3,80 1,37 p = 0,043 12,60 2,37 p = 0,218 l -THP 0,03mg/kg 8 23,37 16,59 p = 0,122 1,00 0,38 p = 0,274 8,50 1,20 p = 0,965 l -THP 0,1mg/kg 9 40,11 13,32 p = 0,017 2,33 0,78 p = 0,035 8,56 1,39 p = 0,968 l -THP 0,3kg/mg 7 29,86 11,19 p = 0,033 1,14 0,40 p = 0,161 7,43 1,46 p = 0,601 l -THP 1mg/kg 9 32,56 14,09 p = 0,035 1,33 0,44 p = 0,133 8,67 0,82 p = 0,905 l -THP 3mg/kg 9 18,00 7,25 p = 0,133 1,56 0,53 p = 0,211 9,33 1,53 p = 0,905 l -THP 10mg/kg 9 24,56 7,55 p = 0,010 1,56 0,44 p = 0,053 9,44 1,31 p = 0,720 l -THP 100mg/kg 10 57,20 28,70 p = 0,075 1,00 0,39 p = 0,280 3,40 0,91 p = 0,029 Ghi ch. DZP: diazepam; l - THP: l tetrahydropalmatin; gi tr p so snh vi l ung nc mui sinh l cha 1% tween 80. Nhn xt: Diazepam liu 2mg/kg th hin tc dng gii lo u r rt, lm tng c ngha thi gian lu ti tay h, tng s ln vo tay h so vi nhm chng ung nc mui sinh l cha 1% tween 80. l tetrahydropalmatin cc mc liu t 0,1 27 mg/kgn1mg/kgvmcliu10mg/kgthhintcdnggiilourrt,lm tng c ngha thi gian lu li tay h; trong , mc liu 0,1 mg/kg cn lm tng c ngha s ln vo tay h so vi l chng. Trn tng s ln lu li tay kn v tay h,ltetrahydropalmatinmcliu100mg/kglmgimcnghachsny, diazepam v l tetrahydropalmatin cc mc liu cn li khng c s khc bit c ngha so vi l chng.Dovy,tccktquthuctrntisdngmcliul tetrahydropalmatin 0,1 mg/kg, 0,3 mg/kg, 1 mg/kg v 3 mg/kg tin hnh cc th nghim tip theo. 3.1.1.2. Tc dng gii lo u ca l tetrahydropalmatin s dng liu n v liu lp li 5 ngy. Trn cc mc liu la chn ca l- tetrahydropalmatin (t 0,1 mg/kg n 3 mg/kg) thc hin nh gi tc dng gii lo u khi s dng liu n v liu lp lp li 5 ngy trn m hnh EPM. - S dng liu n: Kt qu thu c trn thi gian lu ti tay h; s ln vo tay h v tng thi gian lu li tay kn, tay h ca cc l chut th nghim trong m hnh ch thp nng cao c th hin hnh 3.1 v hnh 3.2. Hnh 3.1. Tc dng ca l tetrahydropalmatin v diazepam s dng liu n trn thi lu li tay h, DZP: diazepam, * p < 0,05 so vi l chng ung nc mui sinh l cha 1% tween 80 (n = 9 - 10 con/l). 28 Hnh 3.2. Tc dng ca l tetrahydropalmatin v diazepam s dng liu n trn s ln vo tay h v tng s ln lu li tay kn, tay h; DZP: diazepam, * p < 0,05 so vi l chng ung nc mui sinh l cha 1% tween 80 (n = 9 - 10 con/l). Nhnxt:Sdngliundiazepam2mg/kgvltetrahydropalmatincc mc liu t 0,1 mg/kg n 1 mg/kg th hin tc dng gii lo u r rt, lm tng c ngha thi gian lu ti tay h so vi nhm chng ung nc mui sinh l cha 1% tween 80 (hnh 3.1). S ln vo tay h v tng s ln lu li tay kn, tay h khng c s khc bit c ngha so vi l chng ung nc mui sinh l cha 1% tween 80khisdngliundiazepamvltetrahydropalmatinccmcliula chn (hnh 3.2). - Sau khi ung liu lp li 5 ngy: Kt qu thu c trn thi gian lu ti tay h c trnh by trong hnh 3.3; s ln vo tay h v tng s ln lu li tay kn, tay h ca cc l chut th nghim trong m hnh ch thp nng cao c trnh by trong hnh 3.4. 29 Hnh 3.3. Tc dng ca l tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn thi lu li tay h, DZP: diazepam, * p < 0,05 so vi l chng ung nc mui sinh l cha 1% tween 80 (n = 9 - 10 con/l). Hnh 3.4. Tc dng ca l tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn s ln vo tay h v tng s ln lu li tay kn, tay h; DZP: diazepam, * p < 0,05 so vi l chng ung nc mui sinh l cha 1% tween 80 (n = 9 - 10 con/l). 30 Nhnxt:Sliulpli5ngydiazepamliu2mg/kgvl tetrahydropalmatin cc mc liu t 0,1 mg/kg n 1 mg/kg th hin tc dng gii lo urrt,lmtngcnghathigianlutitayh.Trong,l tetrtahydropalmatin 0,1 mg/kg lm tng c ngha s ln vo tay h, cc mc liu cnlikhngcskhcbitcnghasovilchngungncmuisinhl cha 1% tween 80. Tng s ln lu li tay kn, tay h ca cc l khng c s khc bit c ngha so vi l ung nc mui sinh l c cha 1% tween 80. 3.1.2 Trn m hnh ch T nng cao, ETM Trnccmcliulachncaltetrahydropalmatin(t0,1mg/kgn3 mg/kg), ti tin hnh thc hin thm d tc dng ca l tetrahydropalmatin trn mhnhETMxcnhtcdnggiiloucaltetrahydropalmatintheokiu chng ri lon lo u ton th hay chng ri lon hong s. p ng n trnh nguy him (s cao khi khm ph tay h) lin quan n tc dng chng ri lon lo u ton th, th hin bng thi gian tim tng chut ri tay kn v khm ph tay h ti 3 thi im: thi im ban u (baseline), ln t chut th nht (avoidance 1), lntchutthhai(avoidance2).pngchytrnkhibphinhimmi trng nguy himtrn tay h lin quan n tc dng chng ri lon hong s, th hin bng thi gian tim tng chut ri tay h chui vo tay kn (escape).3.1.2.1. Sau khi s dng liu n l tetrahydropalmatin Thi gian tim tng khm ph ban u (baseline), thi gian tim tng ca p ngntrnh(avoidance1vavoidance2),thigiantimtngpngchytrn (escape)cacclchutthnghimkhisdngliunclomipraminvl tetrahydropalmatin c trnh by trong bng 3.2. 31 Bng 3.2. Tc dng ca l - tetrahydropalmatin v clomipramin s dng liu n trn cc ch tiu nh gi ca m hnh ETM Ghi ch: CLO: clomipramin; l - THP: l tetrahydropalmatin; gi tr p so snh vi l ung nc mui sinh l cha 1% tween 80. Nhnxt:Clomipramin25mg/kgvltetrahydropalmatinccmcliut 0,1mg/kgn3mg/kgkhisdngliunkhngthhintcdnggiilou, khng lm thay i c ngha thi gian tim tng khm ph ti thi im ban u (baseline),thigiantimtngcapntrnh(avoidance1,avoidance2)vca p ng chy trn (escape) so vi l chng ung nc mui sinh l cha 1% tween 80.3.1.2.2. Sau khi s dng liu lp li 21 ngy l tetrahydropalmatinKtqutrnccchtiunhgicamhnhchTnngcaocaccl chutthnghimkhisdngliulpli21ngyclomipraminvl tetrahydropalmatin c trnh by trong bng 3.3. LnBaseline (giy) Avoidance 1 (giy) Avoidance 2 (giy) Escape(giy) NaCl 1067,00 38,84213,00 44,30216,40 42,6045,90 13,56 CLO 25 mg/kg 10 66,00 39,01 p = 0,529 187,50 46,02 p = 0,796 242,20 38,54 p = 0,853 21,60 4,38 p = 0,393 l THP 0,1 mg/kg 10 129,50 43,30 p = 0,971 270,20 29,80 p = 0,529 273,30 26,70 p = 0,436 32,90 16,00 p = 0,353 l THP 0,3 mg/kg 8 13,00 3,45 p = 0,829 197,12 50,34 p = 0.897 236,38 32,07 p = 0,897 20,75 5,70 p = 0,173 l THP 1 mg/kg 9 158,67 48,84 p = 0,356 205,11 47,45 p = 0,968 263,33 32,30 p = 0,780 41,89 32,29 p = 0,079 l THP 3 mg/kg 8 91,25 46,70 p = 0,762 168,25 50,18 p = 0,897 300,00 0,00 p = 0,315 20,88 5,26 p = 0,360 32 Bng 3.3. Tc dng ca l - tetrahydropalmatin v clomipramin s liu lp li 21 ngy trn cc ch tiu nh gi ca m hnh ETM Ghi ch: CLO: clomipramin; l - THP: l tetrahydropalmatin; gi tr p so snh vi l chng ung nc mui sinh l cha 1%tween 80. Nhnxt:Sdngliulpli21ngy,clomipraminliu25mg/kgvl tetrahydropalmatin cc mc liu t 0,1 mg/kg n 3 mg/kg th hin tc dng chng hongs,lmtngcnghathigiantimtngcapngchytrn(escape) nhngkhnglmthayithigiantimtngkhmphtithiimbanu (baseline)cngnhthigiantimtngcapngntrnh(avoidance1, avoidance 2) so vi l chng ung nc mui sinh l cha 1% tween 80. 3.2.nh hng trn hot ng t nhin v khm ph ca l-tetrahydropalmatin Trnccmcliulachncaltetrahydropalmatin(t0,1mg/kgn3 mg/kg),thchinthnghimnhginhhngtrnhotngtnhinvkh nng khm ph trn m hnh mi trng m (open field test, OFT); trong : -nhhngtrnkhnnghotngtnhin(locomotion)thhinthng qua: s ng k chut i qua v s ln ng ln bng 2 chn ca chut. - nh hng trn kh nng khm ph (exploration) th hin thng qua: s ln vo trung tm v thi gian lu li trung tm. LnBaseline (giy) Avoidance 1 (giy) Avoidance 2 (giy) Escape(giy) NaCl 8121,12 52,49168,38 50,60222,38 42,1313,88 2,59 CLO 25 mg/kg 7 218,71 52,48 p = 0,232 144,43 55,13 p = 0,867 260,14 39,86 p = 0,536 101,43 51,39 p = 0,040 l THP 0,1 mg/kg 9 121,00 45,97 p = 0,815 143,56 49,59 p = 0,743 267,33 32,67 p = 0,481 92,00 40,19 p = 0,027l THP 0,3 mg/kg 8 8,00 2,21 p = 0,130 91,62 31,71 p = 0,505 166,75 40,84 p = 0,328 100,25 18,13 p = 0,000 l THP 1 mg/kg 9 144,00 49,63 p = 0,673 259,89 26,53 p = 0,200 277,00 23,00 p = 0,370 50,44 3,29 p = 0,000 l THP 3 mg/kg 8 259,12 35,26 p = 0,130 290,50 9,50 p = 0,161 281,62 15,36 p = 0,574 51,25 8,38 p = 0,002 33 3.2.1. Sau khi s dng liu n l tetrahydropalmatin Kt qu thu c thng qua cc ch tiu nh gi trn m hnh OFT cc l chutthnghimkhisdngliundiazepamvltetrahydropalmatinc trnh by trong bng 3.4. Bng 3.4. Tc dng ca l - tetrahydropalmatin v diazepam s dng liu n trn cc ch tiu nh gi ca m hnh OFT Ghi ch: DZP: diazepam; l - THP: l tetrahydropalmatin; gi tr p so snh vi l chng ung nc mui sinh l cha 1%tween 80. Nhn xt: Diazepam 2 mg/kg v l tetrahydropalmatin cc mc liu t 0,1 mg/kg n 3mg/kgkhi s dng liu nkhngnh hng ti hotng t nhin ca chut, khng lm thay i s ng k chut i qua v s ln ng ln bng 2 chncachutsovilchngungncmuisinhlcha1%tween80.l tetrahydropalmatin mc liu 0,3mg/kg v 1 mg/kg lm gim c ngha s ln vo trung tm. Ccmc liu cn li khng c s khc bit c ngha so vi l chng ung nc mui sinh l cha 1% tween 80. Thi gian lu li trung tm ca cc l khng c s khc bit c ngha so vi l chng ung nc mui sinh l cha 1% tween 80.LnS ng k S ln ng ln bng 2 chn S ln vo trung tm Thi gian lu li trung tm (giy) NaCl 1066,60 12,8215,00 4,565,70 1,0823,00 7,60 DZP 2 mg/kg 9 85,78 21,69 p = 0,661 20,00 4,36 p = 0,356 5,56 1,85 p = 0,549 10,00 3,13 p = 0,079 l THP 0,1 mg/kg 11 77,27 10,92 p = 0,512 12,55 3,87 p = 0,605 6,64 1,06 p = 0,605 23,64 4,40 p = 0,605 l THP 0,3 mg/kg 10 34,50 8,67 p = 0,063 4,90 1,06 p = 0,123 1,60 0,22 p = 0,001 37,00 19,01 p = 0,739 l THP 1 mg/kg 9 38,00 9,93 p = 0,113 9,44 4,13 p = 0,315 1,56 0,44 p = 0,001 86,22 41,94 p = 0,780 l THP 3 mg/kg 10 57,30 12,82 p = 0,684 14,50 4,11 p = 0,971 3,90 1,04 p = 0,190 10,50 2,02 p = 0,123 34 3.2.2. Sau khi s dng liu lp li 5 ngy l tetrahydropalmatin Kt qu trn cc ch tiu nh gi ca m hnh OFT cc l chut th nghim khisdngliulpli5ngydiazepamvltetrahydropalmatinctrnhby trong bng 3.5. Bng 3.5. Tc dng ca l - tetrahydropalmatin v diazepam s dng liu lp li 5 ngy trn cc ch tiu nh gi ca m hnh OFT Ghi ch: DZP: diazepam; l - THP: l tetrahydropalmatin; gi tr p so snh vi l chng ung nc mui sinh l cha 1%tween 80. Nhnxt:Diazepamliu2mg/kgvltetrahydropalmatinccmcliut 0,1mg/kgn3mg/kgkhisdngliulpli5ngykhngnhhngtihot ngtnhincachut,khnglmthayisngkchutiquavsln ng ln bng 2 chn ca chut so vi l chng ung nc mui sinh l cha 1% tween80.Ccmcliunycngkhngnhhngtikhnngkhmphca chut, khng lm thay i c ngha s ln v thi gian lu li trung tm so vi l chng ung nc mui sinh l cha 1% tween 80. 3.3. Tc dng an thn ca l tetrahydropalmatin nh hng ca l tetrahydropalmatin cc mc liu khc nhau v diazepam ln thi gian ng do thiopental c trnh by trong hnh 3.5. LnS ng k S ln ng ln bng 2 chn S ln vo trung tm Thi gian vo trung tm (giy) NaCl 957,00 12,0316,78 4,884,11 1,0111,89 3,27 DZP 2 mg/kg 8 64,12 12,59 p = 0,743 14,75 3,12 p = 0,963 5,50 1,39 p = 0,423 17,50 7,12 p = 0,815 l THP 0,1 mg/kg 11 57,55 13,90 p = 0,941 18,91 4,98 p = 0,824 5,64 1,88 p = 1,000 21,00 5,68 p = 0,370 l THP 0,3 mg/kg 9 80,67 16,67 p = 0,258 30,00 5,93 p = 0,113 7,78 2,09 p = 0,161 20,44 6,12 p = 0,387 l THP 1 mg/kg 10 77,50 10,40 p = 0,182 16,50 3,95 p = 0,905 5,40 1,34 p = 0,497 15,60 4,01 p = 0,604 l THP 3 mg/kg 10 44,20 5,94 p = 0,497 10,20 1,52 p = 0,661 3,00 0,47 p = 0,661 5,50 1,40 p = 0,113 35 Hnh 3.5: nh hng trn thi gian ng do thiopental ca diazepam (DZP) v l - tetrahydropalmatin, ** p < 0,01 so vi nhm chng ung nc mui sinh l cha 1% tween 80 (n = 10 con/l). Nhnxt:Tcdnganthncal-tetrahydropalmatinphthucliu;ccmc liu thp 0,03 mg/kg, 3 mg/kg v mc liu th hin tc dng gii lo u 0,1 mg/kg, 0,3mg/kgv1mg/kg,l-THPkhnglmthayicnghathigianngdo thiopental so vi l chng ung nc mui sinh l cha 1% Tween 80. Trong cng iukinthnghim,diazepam2mg/kgvl-tetrahydropalmatinccmcliu cao hn (10 mg/kg v 100 mg/kg) th hin r tc dng an thn, lm tng c ngha thi gian ng do thiopental so vi l chng ung nc mui sinh l cha 1% tween 80. 3.4. Tc dng chng trm cm ca l tetrahydropalmatin Trnccmcliulachncal-tetrahydropalmatin,tinhnhthnghim nh gi tc dng chng trm cm trn m hnh FST. nh gi tc dng chng trm cm ca thuc thng qua 3 tiu ch: thi gian bt ng, thi gian tro hoc ln, thi gianbi.Trong,trngthibtngcangvtthcnghimphnnhtnh trng "tuyt vng" khi chp nhn bn thn chng vi iu kin th nghim; cc hnh vi tro, ln hay bi th hin trng thi mun thot khi iu kin th nghim. 36 Kt qu thu c trn cc ch tiu nh gi ca cc l chut thc nghim trn m hnh FST c trnh by trong bng 3.6. Bng 3.6. nh hng ca clomipramin, l tetrahydropalmatin thng qua cc ch tiu nh gi trn m hnh bi cng bc (FST). LN Thi gian bt ng (giy) Thi gian bi (giy) Thi gian tro hoc ln (giy) NaCl8150,12 9,26120,88 8,3129,00 6,72 Clomipramin 25 mg/kg 8 113,00 15,11 p = 0,028 125,75 10,10 p = 0,574 61,25 7,84 p = 0,010 l tetrahydropalmatin 0,1 mg/kg 7 102,57 18,16 p = 0,014 178,43 15,41 p = 0,009 19,00 3,81 p = 0,281 l tetrahydropalmatin 0,3 mg/kg 7 93,86 16,21 p = 0,009 167,00 11,77 p = 0,009 39,14 8,95 p = 0,463 l tetrahydropalmatin 1 mg/kg 8 113,00 18,02 p = 0,161 143,88 13,69 p = 0,130 43,12 11,97 p = 0,382 l tetrahydropalmatin 3 mg/kg 8 262,38 8,78 p = 0,000 27,50 5,88 p = 0,000 10,12 3,45 p = 0,028 Ghi ch: gi tr p so snh vi l chng ung nc mui sinh l cha 1%tween 80. Nhnxt:Clomipraminmcliu25mg/kgthhintcdngchngtrm cmrrt,lmtngcnghathigiantrovlncachutvlmgimc nghathigianbtngcachutsovilchngungncmuisinhlcha 1%tween80.l-tetrahydropalmatincngthhintcdngchngtrmcmph thucvoliu.ccmcliuthp0,1mg/kg,0,3mg/kg,l-tetrahydropalmatin lm tng c ngha thi gian bi v gim c ngha thi gian bt ng ca chut so vi l ung nc mui sinh l cha 1% tween 80. Tc dng ny khng cn th hin na khi tng liu ln 1mg/kg. mc liu 3 mg/kg, l - tetrahydropalmatin lm tng c ngha thi gian bt ng, ng thi lm gim c ngha thi gian tro v ln, thi gian bi so vi l chng ung nc mui sinh l cha 1% tween 80.3.5. Bn lun Gii lo u l tc dng dc l thn kinh quan trng c nhiu nghin cu tp trungtipcnnhmtmngundcliuvhotchtchitxuttdcliuc hiu qu,t tc dng khngmongmun, thayth hoc b sung cho cc thuc ha 37 dcangcsdngtrongiutr[66].Ccnghincutrcyvcctc dngdclcaltetrahydropalmatinchyumitpchungvocctcdng nh an thn, gim au, h tr cai nghin....Trn c s , nghin cu ny c thc hin gp phn nh gi v khng nh tc dng gii lo u cng nh mt s tc dng dc l hng thn kinh khc ca l tetrahydropalmatin. 3.5.1. V tc dng gii lo u Kt qu t 2 m hnh dc l thc nghim (m hnh EPM v ETM) cho thy l -tetrahydropalmatinthhintcdnggiilourrtvtcdnggiilou c xc nh l tc dng chng hong s.M hnh ch thp nng cao (EPM), mt trong nhng m hnh ph bin nht s dngtrongsnglchiuqugiiloucathuc.Mhnhdatrntmlcng thng, s hi khng iu kin vi nhng khu vc h v cao, thi thm d, khm ph ca ng vt th nghim vi mi trng mi v l. Mt thuc c tc dng gii lo u s lm gim s s hi vi nhng khu vc h v cao; bn nng thm d, thch khm ph vi mi trng l c th hin thng qua vic lm tng s ln vo cnh tay m v thi gian lu li cnh tay ny, tng s ln di chuyn t tay kn sang tay m [24] [57]. Trn m hnh ny, l THP lm tng c ngha thi gian lu li tay hsovilchngungncmuisinhlcha1%tween80,thhintcdng gii lo u tng t thuc i chiu diazepam (bng 3.1). l -THP c tc dng gii lo u r rt liu thp (t 0,1 mg/kg n 1 mg/kg) v gim dn tc dng khi tng liu (t3mg/kgn100mg/kg),thaythvoltcdnganthn.l tetrahydropalmatinmcliu100mg/kglmgimcnghatngslnluli layknvtayhsovinhmchngungncmuisinhlcha1%tween80, iu ny c th c gii thch do tc dng an thn qu mc lm gim kh nng vn ngtnhincachut[30][51].Sosnhktquvtcdnggiiloutrong nghin cu ny vi nghin cu ca Hong Xue v cng s trn cng m hnh EPM cho thy mc liu bt u th hin tc dng gii lo u ca THP cng l mc liu 0,1 mg/kg. Tuy nhin khong liu c tc dng li c s khc bit gia cc nghin cu. Trong nghin cu ny, khong liu c tc dng l t 0,1 mg/kg n 1 mg/kg trong 38 khi nghin cu ca Hong Xue ch ra khong liu c tc dng i vi l THP l t 0,1 mg/kg n 2,5 mg/kg [62] v i vi dl THP l 0,1 mg/kg n 50 mg/kg [63]. S khc bit ny c th c gii thch do c s khc bit v cch ghi nhn ch tiu nh gi. Trong nghin cu ny, chut c tnh vo 1 cnh tay khi c 4 chn ca chut trong cnh tay . V vy, trong khong thi gian 5 pht s ghi nhn thi gian lu li tay kn, thi gian lu li tay h v thi gian chut vng trung tm. Ngc li, trong nghin cu ca Hong Xue, chut c tnh vo 1 cnh tay khi mt phn c th chut cnh tay [62]. Mt nghin cu ca Leung v cng s li cho kt qukhongliuthhintcdnggiiloucadlTHPl0,5mg/kgn50 mg/kg [39].Lachnccmcliut0,1mg/kgn3mg/kgnhgitiptrnm hnh EPM tc dng gii lo u ca l THP khi s dng liu n v s dng liu lp li5ngy.Ktquchothy,lTHPthhintcdnggiilou,lmtngc nghathigianlulitayhcachutsovilchngtngtdiazepamcc mc liu t 0,1mg/kg n 1 mg/kg khi s dng liu n v s dng liu lp li 5 ngy ( hnh 3.1 n hnh 3.4). Kt qu ny ph hp vi kt qu th nghim thm d liu c tc dng ca l THP, khng nh l THP cc mc liu t 0,1 mg/kg n 1 mg/kg c tc dng gii lo u khi s dng liu lp li 5 ngy, ngoi ra tc dng ny cn c th hin khi s dng liu n l THP. M hnh ch thp nng cao kinh in (EPM) vi ch tiu nh gi chnh da trn s ln vo v thi gian lu li tay m ca ng vt thc nghim c coi l m hnhtrnln,khngchophpphnbitccthloukhcnhau[65].Vvy, GraeffvcngsphttrinmhnhchTnngcao(ETM),citindatrn m hnh EPM kinh in phn bit p ng chy trn (escape) khi ng vt thc nghim c tip xc vi tay m (lin quan n ri lon hong s) v p ng n trnhcch(inhibitoryavoidance)saughinhphinhimvivngnguyhim (linquannrilonloutonth)pdngnhgitcdnggiilouca thuc. Theo , s hi c iu kin (conditioned fear) hay p ng n trnh c chctnhlthigianchutritaykntrong3thnghimlintip(avoidance 39 1,2,3), cn s hi khng iu kin (unconditioned fear) l khong thi gian chut trnthotkhitaymhaypngchytrn.TheoDeakinvGraeff,shic iukinclinquannrilonloulanta(GeneralizedAnxietyDisorder-GAD) ngi, trong khi s hi khng iu lin lin quan n ri lon hong s PanicDisorder(PD)[29].Vicchongvtthcnghimtipxcvimhnh (trong vng 30 pht) 24 h trc khi tin hnh th nghim (pre test) gip gim thi gian chy trn do tng nhy pht hin tc dng chng hong s ca cc thuc c thc hin trong nghin cu [55]. Trn m hnh ny, s dng liu lp li 21 ngy, l THP cc mc liu 0,1 mg/kg n 1 mg/kg lm tng c ngha thi gian tim tng ca p ng chy trn (escape) khi chut c t tay m, th hin tc dngchngrilonhongstngtthucichngchngtrmcm3vng clomipramin[50],mkhnglmthayipngntrnh(avoidance)sovil chng (bng 3.2). Trong khi , sau khi sdng liu n, l- THP cc mc liu ny v clomipramin c xu hng lm tng p ng n trnh cng nh gim p ng chytrncachutsovilchng(khngcnghathngk)(bng3.3).Cc bng chng lm sng cho thy rng vic s dng cc thuc chng trm cm ba vng hoc thuc tng t nhng liu u tin lm gia tng s lo lng, sau tc dng gii lo u s xut hin dn khi s dng thuc ko di [40]. Tc dng gii lo u ca tetrahydropalmatin bi khng bi flumazenil,mt cht cnh tranh v tr gn ca benzodiazepin trn receptor GABAA, chng t c ch gii lo u ca THP thng qua, t nhtmt phn, tng tc viv trgn ca benzodiazepin trn receptor GABAA [36][39].giithchchotcdngchnghongscalTHP,ngoicch thngquareceptorGABAA,cchtcdngkhctngtccthuctcdng thng qua h serotonergic cng c cp [48]. 3.5.2. V nh hng trn hot ng t nhin v kh nng khm ph Mhnhmitrngm(OFT)lmhnhnhgihotngtnhin,kh nngkhmphvtnhtrnglolngcachutkhictvomtmitrng mi khc bit hon ton mi trng sng by n [58]. Sau nhiu nghin cu, m hnhOFTccitinnhmphhpvitngiukinthnghim,phhp 40 vimcchcatngnhnghincu[56].Sngkchutiquavsln ng ln bng 2 chn l nhng tiu ch gip nh gi kh nng hot ng t nhin ca chut, khi cc ch s ny tng ln cho thy s gia tng v kh nng hot ng t nhincachut[49].Vngtrungtmlvngnmchnhgia,ckhongkhng gian xung quanh khc bit vi vng vin ngoi gip vi cc bc tng ca m hnh [25]. S ln vo trung tm v thi gian lu li trung tm l nhng tiu ch gip nh gi kh nng khm ph v tnh trng lo lng ca chut, khi cc ch s ny tng cho thy s gia tng v kh nng khm ph v gim tnh trng lo lng ca chut khi tip xc vi mi trng mi [49]. Ngoi cc tiu ch quan st nh hot ng ngm vut (grooming) [32], s lng phn v nc tiu chut[37] cng gip nh gi tnh trng lo lng ca chut. Trn m hnh ny, l - THP cc mc liu gii lo u khng nh hng ti hot ng t nhin ca chut c khi s dng liu n v s dng liu lpli5ngy,khnglmthayicnghasngkchutiquavsln ng ln bng 2 chn ca chut (bng 3.4 v 3.5). Kt qu ny hon ton ph hp vi ktqu v ch s tng s ln lu li taykn v tayh trnmhnh EPM. Kt qu ny cng hon ton ph hp vi cc nghin cu trc , theo nghin cu ca Kai Yue v cng s cho thy l THP mc liu 5 mg/kg tim mng bng (i.p) tnh theo th trng chut lm gim r rt kh nng hot ng trn chut cng, trong khiccmcliuthphnkhngghinhncnhhngny[64].Cngtheo HongXuevcngs,mcliulmgimhotngtnhincadlTHPvlTHPtngngl:t0,1mg/kgn100mg/kgvt5mg/kgn40mg/kgs dng ng ung tnh theo trng lng c th chut [62] [63]. Trn kh nng khm ph ca chut, l THP cc mc liu 0,3 mg/kg v 1 mg/kg lm gim c ngha s ln vo trung tm so vi l chng khi s dng liu n (bng 3.4); trong khi cc mc liu cn li cng nh khi s dng liu lp li 5 ngy khng lm thay i c nghaslnvthigianlulivngtrungtmcachut(bng3.5).Ccbng chng lm sng cho thy rng vic s dng cc thuc chng trm cm ba vng hoc thuc tng t nhng liu u tin lm gia tng s lo lng, sau tc dng gii lo u s xut hin dn khi s dng thuc ko di [40]. 41 3.5.3. V tc dng an thn Ko di thi gian ng do thuc ng barbituric l th nghim kinh in thm d tc dng an thn ca thuc da trn nguyn tc hip ng tc dng ca thuc [45]. Kt qu nghin cu cho thy l THP mc liu thp (t 0,03 mg/kg n 3 mg/kg) khngthhintcdnganthn;tuynhinmcliucaohn(10mg/kgv100 mg/kg) l THP th hin tc dng an thn r rt tng t diazepam, lm ko di c ngha thi gian ng do thiopental so vi l chng (hnh 3.5). Kt qu nghin cu nygipkhngnhtcdnganthncalTHPnhngmcliuthphnso vi cc nghin cu trc y cho thy tm quan trng ca vic la chn liu c tc dng.NguynLanThyTyvcngsquanstthyStilux-60(l-tetrahydropalmatin)thhintcdngkodithigianngdothiopentalgp khong7lnkhidngliu48mg/kgtrnchutnhttrng[16].TheoPhmDuy Mai v cng s, gindarin hydrochlorid (l tetrahydropalmatin hydrochlorid) mc liu 30 mg/kg th trng chut ko di thi gian ng do thiopental ln 1,5 ln so vi lchng[12].TheoNguynTinVng,lTHPmcliu40mg/kgthtrng chut lmtng thi gian ng dothiopental ln 8 ln so vi nhmchng [18]. Tc dnganthncatetrahydropalmatincchngminhclinquannkh nng hot ha receptor GABAA [36] v lin quan ti tc dng i khng ca thuc trn receptor D1, D2 v D3 [38]. 3.5.4. V tc dng chng trm cm Lo u, trm cm, an thn v c ch sinh ha dn truyn thn kinh c s tng ng, u thng qua cc h thng dn truyn thn kinh v cht dn truyn thn kinh nhnoradrenergic,seretogenic,gamma-aminobutyric-acid(GABA)vserotonin (5-HT)[23].iunybiuhinccbnhnhnbrilonhongsthngc triu chng trm cm km theo hoc khng km theo ri lon trm cm v cc bnh nhn trm cm c th gp cn hong lon khi qu lo lng, ni cch khc, ri lon lo u v trm cm thng i km vi nhau [33]. Cc thuc iu tr ri lon lo u cng thng c tc dng chng trm cm v ngc li nh: nhm thuc c ch chn lc thu hi serotonin (SSRI) i din l fluoxetin, paroxetin, setralin; nhm thuc chng 42 trmcmbavng(TCAs)cidinlimipramin,clomipramin...[4].Vvy, nghin cu tip tc tin hnh nh gi tcdng chng trm cm trnm hnh bi cng bc (forced swim test, FST) cal THP ti cc mc liu th hin tc dng gii lo u. M hnh FST c pht trin bi Porsolt v cng s vo nm 1977 v s dng rng ri trong nghin cu tin lm sng nh gi tc dng chng trm cm ca cc thuc [34]. M hnh ny sau c ci tin nhm khc phc nhc im v tng nhy pht hin tc dng chng trm cm ca thuc, cho php nh nghin cu phn bit c cc hnh vi c th trong hot ng ca ng vt th nghim nh: bi, tro, bt ng [28]. c bit, m hnh FST ci tin ghi nhn cc thuc lin quan ti h noradrenergic lm gim trng thi bt ng km theo s tng tng ng ca hnh vi tro, trong khi cc thuc lin quan ti serotonin nh SSRIs lm gim trng thi bt ng nhng tng hnh vi bi ca ng vt thc nghim [28] [31]. Kt qu nghincuchothylTHPccmcliu0,1mg/kgv0,3mg/kgthhintc dngchngtrmcmtuynhinkhngrrtbngclomipramin,cth: clomipraminvlTHPulmgimcnghathigianbtngcachut, ngthiclomipraminlmtngcnghathigiantro/ln(hnhngctnh cht mnh m hn) trong khi l THP lm tng c ngha thi gian bi ca chut so vi l chng (bng 3.6). l THP mc liu 3 mg/kg lm tng c ngha thi gian btng,gimcnghathigiantro/lnvthigianbicachutsovil chng ung nc mui sinh l. iu ny c th c gii thch do tc dng an thn ca l THP lm gim cc hot ng vn ng ca chut. Nh vy, mc liu th hin tc dng gii lo u (0,1 mg/kg v 0,3 mg/kg), l THP th hin tc dng chng trm cm; iu ny hng ti gi thuyt v c ch tc dng ca l - THP c th lin quantidntruynserotonintngtcchchngrilonhongsvchng trmcmcaccthucchngtrmcm3vnghaycchchnlctithuhi serotonin [28]. Nhvy,ktqucanghincuvtcdnggiiloucalTHPcs trng lp tng i so vi cc nghin cu trc , khng nh tc dng gii lo u ca l THP vmc liu th hin tc dng t 0,1 mg/kg n 1 mg/kg (theo nghin 43 cucaHongXuevcngsmcliuthhintcdngt0,1mg/kgn2,5 mg/kg)[62].TcdnggiiloucalTHPcxcnhltcdngchng hong s thng quam hnh ch T nng cao khi s dng liu lp li 21 ngy. Ti ccmcliuthhintcdnggiilou,lTHPkhnglmnhhngtihot ng t nhin vkhnng khm ph ca chut trnm hnhmi trngm, kt qunyphhpviccnghincutrccaKaiYue[64],nghincuca Hong Xue [62]. ng thi, ti cc mc liu ny, l THP khng th hin tc dng an thn v th hin tc dng chng trm cm; tc dng an thn ch xut hin cc mc liu cao hn ( 10 mg/kg v 100 mg/kg) v mc liu ny thp hn so vi cc nghin cu trc ca Nguyn LanThuyTy, nghin cu caNguyn Tin Vng...cho thytmquantrngcavicchnliuctcdng.Ccktqunghincunh hngcchtcdngcalTHPkhngchlinquantikhnnghotha receptor GABAA v hot ng i khng ca thuc trn receptor D1, D2, D3 m cn hng ti gi thuyt v c ch tc dng lin quan ti dn truyn serotonin tng t c ch tc dng ca cc thuc chng trm cm 3 vng hay c ch chn lc ti thu hi serotonin [28] [48]. 44 KT LUN V XUT KT LUN Tktquthuctrnccmhnhdclthcnghimcthchin trong ti chng ti rt ra mt s kt lun sau: 1.l tetrahydropalmatin (l THP) khi s dng liu n v liu lp li 5 ngy ticcmcliut0,1mg/kgn1mg/kgsdngngungthhintc dnggiiloutrnmhnhchthpnngcaotngtchtichiu diazepam2mg/kg.TcdnggiiloucxcnhtrnmhnhchT nngcaoltcdngchngrilonhongstngtchtichiu clomipramin 25 mg/kg khi s dng liu lp li 21 ngy v tc dng ny khng xut hin khi s dng liu n l - THP. 2.Ti cc mc liu th hin tc dng gii lo u (0,1 mg/kg n 1 mg/kg), l-tetrahydropalmatin khng lm nh hng ti kh nng hot ng t nhin, kh nng khm ph ca chut (m hnh mi trng m) ng thi khng th hin tcdnganthn(mhnhtngcngthigiandothiopental).Ticcmc liuny,ltetrahydropalmatinthhintcdngchngtrmcm(mhnh bi cng bc) tng t cht i chiu clomipramin liu 25 mg/kg. XUT 1.Nghincutcdnggiiloucaltetrahydropalmatintrnccm hnh dc l thc nghim i vi: ng vt chu stress do c lp, ng vt chustressdosuygimhormonsinhdc(chutcisaukhictbung trng)... 2.Nghin cu c ch dc l phn t lin quan n tc dng gii lo u ca l tetrahydropalmatin theo hng dn truyn serotoninergic TI LIU THAM KHO TI LIU THAM KHO TING VIT 1.DngPhcAn,HunhThBchHuyn,NguynNgcKhi(2009), "Kho st hot tnh gii lo u ca mt stinh du tvqucyCitrush Rutaceace", Tp ch Dc hc. 10, tr. 49-53. 2.Bi Th Bng v cng s (1986), "Kho st hm lng l - tetrahydropalmatin trong c bnh vi mc hoang Vit Nam", Cng trnh nghin cu khoa hc Vin Dc Liu 1972 - 1986, NXB Y hc, H Ni, tr. 50 - 52. 3.B Y t (2002), Dc Th Quc Gia Vit Nam, NXB Y hc, tr. 291, 369. 4.B Y t (2007), Dc l hc tp I, NXB Y hc, H Ni, tr. 140 - 146. 5.B Y t (2007), K thut sn xut dc phm, tp I, NXB Y hc, H Ni, tr. 228. 6.B Y t (2009), Dc in Vit Nam IV, NXB Y hc7.PhmThDuyn(2004),nhlngL-tetrahydropalmatintrongmts loi Bnh vi Vit Nam v hai ch phm vin nang cng Heantos 4 v vin hon Balok bng phng php sc k lng hiu nng cao (HPLC), Lun n Thc s Dc hc, i hc Dc H Ni. 8.PhmThDuyn,TrnhVnLu,PhmThanhK(2003),nhlngL- tetrahydropalmatin(Rotundin)trongdcliubnhvibngphngphp sc k lng hiu nng cao (HPLC), Thng bo kim nghim, H Ni. 9.Trungm,ThPhng(2006),"Tcdnganthncasenin,bt alcaloid l sen", Tp ch Dc Hc. 368, tr. 19 - 22. 10.V Xun Giang (2003), Nghin cu loi Stephania viridiflavens H.S.Lo et M. Yang thu hi Sn La, Lun n Thc s Dc hc, i hc Dc H Ni. 11.NguynQucHuy(2010),Nghincuvthcvt,thnhphnhahc, mtstcdngsinhhccamtsloithucchiStephaniaLour. Vit Nam, Lun n Tin s Dc hc, i hc Dc H Ni. 12.Phm DuyMai, Phanc Thun (1986), "Tc dng dc l cabnh vi", Cng trnh NCKH Vin Dc liu 1972 - 1986, NXB Y hc, H Ni, tr. 55 - 57. 13.LnhMi,TrnMinhHi,DngcHuyn,TrnHuyThi,Ninh KhcBn (2005), TinguynthcvtVitNam(nhngcychacc hot cht c hot tnh sinh hc), tp I. 14.Ng i Quang (1999), Tp ch cng nghip ha cht, NXB Vinachem. 15.NguynMinhTun(2004),Ccrilontmthn:Chnonviutr, NXB Y hc, H Ni. 16.Nguyn Lan Thy Ty, V Phng Nguyn, NguynHuc (2008), "nh gi tc dng an thn-gy ngca 2 chphm c ngun gc dc liu", Tp ch Y hc Thnh ph H Ch Minh. 12, tr. 108 - 112. 17.Vin Dc Liu (2004), Cy thuc v ng vt lm thuc Vit Nam, tp 1, NXB Khoa hc k thut, H Ni. 18.Nguyn Tin Vng (2000), Nghin cu v thc vt, ha hc v tc dng sinh hc ca mt s loi thuc chi Stephania Lour. Vit Nam, Lun n Tin s Dc hc, i hc Dc H Ni. 19.PhmVnVng,NguynThBchLuyn(2007),"Tcdngdclca siro Laroxen len mt s chc nng thn kinh trung ng trn thc nghim", Tp ch Y Dc hc Qun s. 1, tr. 4 - 7. TI LIU THAM KHO TING ANH 20.American Psychiatric Association (2000), Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. 21.AnderssonC.,BergarpE.,HedmanG.(1992),"Sick-listedbutactive", Lakartidningen. 89(5), pp. 281 - 283. 22.BandessariniR.J.(2006),"DrugTherapyofDepressionandAnxiety",in BrutonL.L.,LazoJ.S.,ParkerK.L.,Editor,Goodman&Gilmans:The PharmacologicalBasisofTherapeutics,McGraw-HillCompanies,New York, pp. 429 - 459. 23.BuschmannH.,DiazJ.J.,HolenzJ.,PrragaA.,TorrensA.andVelaJ.M. (2007),Antidepressants,Antipsychotics,Anxiolytics,Wiley-VCHVerlag GmbH & Co. 24.CarobrezA.,BertoglioL.(2005),"Ethologicalandtemporalanalysesof anxiety-likebehaviour:Theelevatedplus-mazemodel20yearson", Neurosci. Biobehav. Rev. 29, pp. 1193 - 1205. 25.CarreyN.,McFadyenM.P.,BrownR.E.(2000),"Effectsofsubchronic methylphenidatehydrochlorideadministrationonthelocomotorand exploratorybehaviorofprepubertalmice",JChildAdolesc Psychopharmacol. 10(4), pp. 277 - 286. 26.ChuH.Y.,JinG.Z.,EtianF.,ZhenX.C.(2008),"Recentdevelopmentin studies of tetrahydroprotoberberines: Mechanism in antinociception and drug addition", Cell Mol Neurobiol. 28, pp. 491 - 499. 27.ChuehF.Y.,HsiehM.T.,ChenC.F.,LinM.T.(1995),"Hypotensiveand bradycardiceffectsofdl-tetrahydropalmatinemediatedbydecreasein hypothalamicserotoninreleaseintherat",Jpn.J.Pharmacol.69,pp.177- 180. 28.Cryan J.F., Markou A., Lucki I. (2002), "Assessing antidepressant activity in rodents:recentdevelopmentsandfutureneeds",TRENDSin Pharmacological Sciences. 23(5), pp. 238 - 245. 29.DeakinJ.F.W.,GraeffF.G.(1991),"5-HTandmechanismsofdefence",J. Psycho-pharmacol. 5, pp. 305 - 315. 30.Dowson G., Tricklebank M. (1995), "Use of the evlevated plusmaze in the search for novel anxiolytic agents", Trends Pharmacol. Sci. 16, pp. 33 - 36. 31.DumanC.H.(2010),"ModelsofDepression",inLitwackG.,Editor, Vitamins and Hormones, Academic Press, pp. 1 - 21. 32.EspejoE.F.(1997),"Effectsofweeklyordailyexposuretotheelevated plus-maze in male mice. ", Behav Brain Res. 87(2), pp. 233 - 238. 33.FainmanD.(2004),"Examiningtherelationshipbetweenanxietydisorders anddepression",CanadianMedicalEducationJournal.22(10),pp.568- 571. 34.GohilK.J.,LawarM.A.(2012),"RodentAntidepressantModelsin Neuropsychopharmacology:ALaboratoryPerspective",International Journal of Research in Pharmacy and Science. 2(1), pp. 21 - 43. 35.GraeffF.G.,NettoC.F.,JrH.l.Z.(1998),"TheelevatedT-mazeasan experimentalmodelofanxiety",NeuroscienceandBiobehavioral Reviews. 23, pp. 237 246. 36.HalbsguthC.,MeibnerO.,HaberleinH.(2003),"Positivecooperationof protoberberinetype2alkaloidsfromCorydaliscavaontheGABAA binding site", Planta Med. 69, pp. 305 - 309. 37.Hall C.S. (1934), "Emotional behavior in the rat 1. Defecation and urination as measures of individual differences in emotionality", J. Comp. Psychol. 18, pp. 382 - 403. 38.JinG.Z.,XuS.X.,YuL.P.(1986),"Differenteffectsofenantiomersof tetrahydropalmatineondopaminergicsystem",Scient.Sin.Ser.B.Chem. Biol., Agric., Med. Earth Sci. 29(10), pp. 1054 - 1064. 39.LeungW.C.,ZhengH.,HuenM.,LawS.L.,XueH.(2003),"Anxiolyticlike actionoforallyadministereddl-tetrahydropalmatineinelevatedplusmaze", ProgressinNeuro-Psychopharmacology&BiologicalPsychiatry.27,pp. 775 779. 40.Liebowitz (1989), "Antidepressants in panic disorders", Psychiatry. 155, pp. 46 - 52. 41.ListerR.G. (1987), "Theuseofaplus-mazetomeasureanxietyinthe mouse", Psychopharmacol. 92, pp. 180 - 185. 42.LiuX.,YangZ.,LiR.,XieJ.,YinQ.,BloomA.S.,LiS.-J.(2012), "Responsesofdopaminergic,serotonergicandnoradrenergicnetworksto acutelevo-tetrahydropalmatineadministrationinnaverats detectedat 9.4 T", Magn Reson Imagin. 30(2), pp. 261 - 270. 43.LiuG.Q.,AlgeriS.,GarattiniS.(1982),"dl-Tetrahydropalmatineas monoamine depletor", Arch. Int. Pharmacodyn. Ther. 258(1), pp. 39 -50. 44.LiuY.-L.,YanL.-D.,ZhouP.-L.,WuC.-F.,GongZ.-H.(2009), "Levotetrahydropalmatineattenuatesoxycodone-inducedconditionedplace preference in rats", European Journal of Pharmacology. 602, pp. 321 - 327. 45.MaY.,HanH.,EunJ.S.,KimH.C.,HongJ.T.,OhK.W.(2007), "SanjoinineAisolatedfromZizyphiSpinosisemenaugments pentobarbital-inducedsleepingbehaviorsthroughthemodificationof GABA-ergic systems", Biol. Pharm. Bull. 30, pp. 1748 - 1753. 46.MaZJ.,LiXD.,GuXZ.,ChengLP.,MaoSJ.,(2006),"Effectsof different types and standard of processing vinegaron inherent constituents in rhizoma of Corydalis Yanhusuo", Zhongguo Zhong Yao Za Zhi 31(6), pp. 465 - 467. 47.Mantsch J.R., LiS.-J., Risinger R., Awad S., KatzE.,Baker D.A.,Yang Z. (2007),"Levo-tetrahydropalmatineattenuatescocaineselfadministrationandcocaine-inducedreinstatementinrats", Psychopharmacology. 192, pp. 581 - 591. 48.Pinheiro S., Zangrossi H., Del-Ben C.,Graeff F.G. (2007), "Elevated maze asanimalmodelsofanxiety:effectsofserotonergicagents",Anaisda Academia Brasileira de Ciencias. 79, pp. 71 - 85. 49.PodhornaJ.,BrownR.E.(2002),"Straindifferencesinactivityand emotionalitydonotaccountfordifferencesinlearningandmemory performance between C57BL/6 and DBA/2 mice", Genes Brain Behav. 1(2), pp. 96 - 110. 50.Poltronieri S., Zangrossi H., Viana M.d.B., (2003), "Antipanic-like effect ofserotoninreuptakeinhibitorsintheelevatedT-maze", Behav.Brain Res. 147, pp. 185 - 192. 51.Rodgers R.,Cao B.-J., Dalvi A.,Holmes A. (1997), "Animal modelsof anxiety: an ethological perspective", Brazilian J. Med. Biol. Res. 30, pp. 299 - 304. 52.SarrisJ.,PanossianA.,SchweitzerI.(2011),"Herbalmedicinefor depression,anxietyandinsomnia:Areviewofpsychopharmacologyand clinical evidence", Eur. Neuropsychol. 21, pp. 841-860. 53.SlattleyD.A.,CryanJ.F.(2012),"Usingtheratforcedswimtesttoassess antidepressant-likeactivityinrodents",Natureprotocol.7(6),pp.1009 1014. 54.SutinE.L.,JacobowitzD.M.(1991),"Neurochemicalsinthedorsalpontine tegmentum", Prog. Brain Res. 88, pp. 3 - 14. 55.TeixeiraR.C.,Jr.H.Z.,GraeffF.G.(2000),"BehavioralEffectsofAcute andChronicImipramineintheElevatedT-Maze ModelofAnxiety", Pharmacology Biochemistry and Behavior. 65(4), pp. 571 - 576. 56.ToddD.G.,DavidT.D.,ColleenE.K.,(2009),"Theopenfieldtest",in Todd D.G., Editor, Mood and Anxiety Related Phenotypes in Mice, Humana Press, pp. 1 - 20. 57.Walf A.A., Frye C.A. (2007), "The use of the elevated plus maze as an assay of anxiety related behavior in rodents", Nature protocols. 2, pp. 322 - 328. 58.Walsh R.N., Cummins R.A. (1976), "The Open-Field Test: a critical review", Psychol Bull. . 88(3), pp. 482 - 504. 59.WangJ.B., MantschJ.R. (2012), "L-tetrahydropalamatine:apotential newmedicationforthetreatmentofcocaine addiction", FutureMed. Chem. 4(2), pp. 177 - 186. 60.XiZ.X.,GilbertJ.,CamposA.C.,KlineN.,AshbyC.R.Jr,HaganJ.J., HeidbrederC.A.,GardnerE.L.(2004),"Blockadeofmesolimbicdopamine D3 receptors inhibits stress-induced reinstatement of cocaineseeking in rats", Psychopharmacology. 176, pp. 57 - 65. 61.XuS.X.,JinG.Z.,YuL.P.,LiuG.X.,LuW.W.,FangS.D.(1987),"Brain dopamine depleted by d-tetrahydropalmatine", Acta Pharm. Sin. 8, pp. 207 - 212. 62.XueH.(2004),usesforltetrahydropalmatin,PatentApplication Publication, Editor, United States, Pub.No. US 2004/0198770 A1 pp. 1 - 6. 63.XueH.,LawS.L.(2002),NovelusesofdlTHP,PatentApplication Publication, Editor, United States, Pub. No. US 2002/0143067 A1, pp. 1 - 6. 64.Yue K., Ma B., Ru Q., Chen L., Gan Y., Wang D., Jin G.,Li C. (2012), "Thedopaminereceptorantagonistlevo-tetrahydropalmatineattenuatesheroinself-administrationandheroin-inducedreinstatementinrats.", Pharmacology, Biochemistry and Behavior. 102, pp. 1 - 5. 65.ZangrossiH.,GraeffF.(1997),"Behavioralvalidationoftheelevated Tmaze, a new animal model for anxiety", Brain Res. Bull. 44, pp. 1 - 5. 66.ZhangZ.(2004),"Therapeuticeffectsofherbalextractsandconstituentsin animal models of psychiatric disorders", Life Sci. 75, pp. 1659 - 1669.