Research Article Protective Effect of Artemisia asiatica...

Research ArticleProtective Effect of Artemisia asiatica Extract and Its ActiveCompound Eupatilin against Cisplatin-Induced Renal Damage

Jun Yeon Park1 Dahae Lee1 Hyuk-Jai Jang2 Dae Sik Jang3 Hak Cheol Kwon4

Ki Hyun Kim5 Su-Nam Kim4 Gwi Seo Hwang1 Ki Sung Kang1 and Dae-Woon Eom6

1College of Korean Medicine Gachon University Seongnam 461-701 Republic of Korea2Department of Surgery University of Ulsan College of Medicine Gangneung Asan Hospital Gangneung 210-711 Republic of Korea3Department of Life and Nanopharmaceutical Science College of Pharmacy Kyung Hee University Seoul 130-701 Republic of Korea4Natural Products Research Center Korea Institute of Science and Technology Gangneung 210-340 Republic of Korea5School of Pharmacy Sungkyunkwan University Suwon 440-746 Republic of Korea6Department of Pathology University of Ulsan College of Medicine Gangneung Asan Hospital Gangneung 210-711 Republic of Korea

Correspondence should be addressed to Ki Sung Kang kkanggachonackr and Dae-Woon Eom edwjyhgnahcokr

Received 13 April 2015 Accepted 14 June 2015

Academic Editor Avni Sali

Copyright copy 2015 Jun Yeon Park et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The present study investigated the renoprotective effect of anArtemisia asiatica extract and eupatilin in kidney epithelial (LLC-PK1)cells Although cisplatin is effective against several cancers its use is limited due to severe nephrotoxicity Eupatilin is a flavonoidcompound isolated from the Artemisia plant and possesses antioxidant as well as potent anticancer properties In the LLC-PK1cellular model the decline in cell viability induced by oxidative stress such as that induced by cisplatin was significantly and dose-dependently inhibited by the A asiatica extract and eupatilin The increased protein expressions of phosphorylated JNK and p38by cisplatin in cells were markedly reduced after A asiatica extract or eupatilin cotreatment The elevated expression of cleavedcaspase-3 was significantly reduced byA asiatica extract and eupatilin and the elevated percentage of apoptotic cells after cisplatintreatment in LLC-PK1 cells wasmarkedly decreased by cotreatmentwithA asiatica extract or eupatilin Taken together these resultssuggest that A asiatica extract and eupatilin could cure or prevent cisplatin-induced renal toxicity without any adverse effect thusit can be used in combination with cisplatin to prevent nephrotoxicity

1 Introduction

Cisplatin is a potent chemotherapeutic agent for the treat-ment of multiple human malignancies [1 2] It accumulatesin all segments of nephron but is predominantly taken up bythe proximal tubule cells which then provokes severe damage[3] The efficacy of cisplatin is dose dependent but the sideeffect in kidney limits the use of higher doses to improve itschemotherapeutic effects [4 5] The toxic effects of cisplatinmainly occur via oxidative stress and DNA damage [6 7]ultimately leading to apoptotic pathways in tumour cells [8]and also in renal cells [4 9 10]

For centuriesmany natural products have been identifiedfor the prevention andor treatment of kidney diseasesbecause they are believed to have nephroprotective effects

They are widely used in clinical practice in many partsof the world For example Silybum marianum was foundto attenuate nephrotoxicity induced by gentamicin in dogs[11] A water extract of Kalanchoe pinnata leaves protectedrat kidneys from gentamicin-induced nephrotoxicity [12]Salviae Radix extract exerted a protective effect againstcisplatin-induced renal cell injury and its effect might bemediated by its antioxidant effect [13]

Artemisia asiaticaNakai is a traditional oriental medicineand it has been used for the treatment of several inflam-matory disorders Recent studies revealed that A asiaticahas antioxidative and anti-inflammatory effects contributingto its protective effects against various pathophysiologicalconditions including gastric damage [14] liver damage [15]experimental pancreatitis [16] and tumor promotion [17]

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2015 Article ID 483980 6 pageshttpdxdoiorg1011552015483980

2 Evidence-Based Complementary and Alternative Medicine

HO

H3CO

OOH

OCH3

OCH3

O

(a)

0

20

40

60

80

100

120

0 10 25 50 100

DPP

H ra

dica

l sca

veng

ing

activ

ity

Vitamin CArtemisiaEupatilin

Concentration (120583gmL)

(b)

120

100

80

60

40

20

0

0

10

25

50

100

250

Cisplatin (25120583M)

Con

trol

Cel

l via

bilit

y (

of c

ontro

l)

Artemisia (120583gmL)

(c)

120

100

80

60

40

20

0

0

10

25

50

100


Con

trol

Cel

l via

bilit

y (

of c

ontro

l)

Eupatilin (120583gmL)

(d)

Figure 1 Effects of A asiatica extract and eupatilin on cisplatin-induced nephrotoxicity in LLC-PK1 cells (a) Structure of eupatilin(b) Comparison of DPPH radical scavenging effects ofA asiatica extract eupatilin and vitamin C (c) Dose-dependent protective effect ofAasiatica extract against cisplatin-induced nephrotoxicity in cells (d) Dose-dependent protective effect of eupatilin against cisplatin-inducednephrotoxicity in cells

Stillen is a commercially available extract fromA asiaticaEupatilin (Figure 1(a)) an active compound isolated fromA asiatica has been reported to treat peptic ulcers andgastritis It has antioxidative and anti-inflammatory effects

against gastric mucosal injury [18 19] Various inflammatorymediators such as cytokines and oxidative stress that canaffect gastric mucosal injury are thought to be involved inits action mechanism [20 21] Eupatilin was also reported to

Evidence-Based Complementary and Alternative Medicine 3

have therapeutic potential for the treatment of gastric cancer[22 23]

Although cisplatin-induced nephrotoxicity has been welldocumented the effects of A asiatica and eupatilin on apop-tosis in kidney cells after cisplatin exposure remain underactive investigation

2 Materials and Methods

21 Chemicals and Reagents An ethanolic extract of Aasiatica and its active compound eupatilin were preparedas reported previously [17 18] Cisplatin and 11-diphenyl-2-picryl-hydrazyl (DPPH) were purchased from Sigma Chemi-cal Co (St Louis MO USA)The stock solution of chemicalswas prepared in 100 dimethylsulfoxide (DMSO) and storedat minus20∘C until use Antibodies for p38 p-p38 JNK p-JNKERK p-ERK cleaved caspase-3 andGAPDHwere purchasedfrom Cell Signaling (Boston MA USA)

22 Protective Effect against Cisplatin-Induced Nephrotoxicityin Cells Possible renoprotective effects against cisplatin-induced damage were evaluated in LLC-PK1 cells as reportedpreviously [24] In brief LLC-PK1 cells were seeded in 96-well culture plates at 1 times 104 cells per well and the test sampleandor radical donor 25 120583M cisplatin were added to theculture medium Twenty-four hours later the cell viabilitywas measured by using a microplate reader (PowerWave XSBio-Tek Instruments Winooski VT USA)

23 DPPH Radical Scavenging Assay The radical scavengingactivity of A asiatica and eupatilin against DPPH was deter-mined spectrophotometrically In microwells 100120583L of anaqueous solution of the completely dissolved sample (control100 120583L DW) was added to an ethanolic solution of DPPH(100 120583L 60 120583M) according to the reported method [25] Thefinal concentrations of the tested samples in the assayedsolution were 10 25 50 and 100 120583gmL Vitamin C was usedas the standard for comparisonThe ability to scavengeDPPHradicals was calculated in terms of percentage of inhibitionaccording to the following equation inhibition = [(119860

0minus

119860

1)119860

0times 100] where 119860

0is the absorbance of the control

(without extract) and 1198601is the absorbance in the presence

of the extract

24 Western Blot Analysis Proteins (whole cell extracts30 120583glane) were separated by electrophoresis in a precast 4ndash15 Mini-PROTEAN TGX gel (Bio-Rad CA USA) blottedonto PVDF transfer membranes as reported previously [26]Bound antibodies were visualized using ECL Advance West-ern Blotting Detection Reagents (GE Healthcare UK) and aLAS 4000 imaging system (Fujifilm Japan)

25 Image-BasedCytometric Assay Todetermine the portionof the population that had become apoptotic cells werestainedwith annexinV-Alexa Fluor 488 conjugate using aTaliimage-based cytometer (Invitrogen CA USA) [27] Propid-ium iodide (PI) was used to differentiate dead cells (annexinV-positivePI positive or annexin V-negativePI positive)

minus

minusminus

+

minusminus

+

minus+

+

+minus

minus minus minus minus

+

minusminus

+

Cleaved caspase-3

p-p38

GAPDH

p38

p-ERK

ERK

p-JNK

JNK

Cisplatin (25120583M)Artemisia (250120583gmL)

Eupatilin (10120583gmL)Eupatilin (50120583gmL)

Figure 2 Involvement of the MAPKs-caspase-3 signaling pathwayin the protective effect of A asiatica extract and eupatilin againstcytotoxicity in cultured LLC-PK1 cells Results of the Western blotshow the levels of p-p38 p38 p-JNK JNK p-ERK ERK andcleaved caspase-3 in LLC-PK1 cells treated with A asiatica extractand eupatilin andor cisplatin at different concentrations for 24 hWhole cell lysates (20120583g) were separated by SDS-PAGE transferredonto PVDF transfer membranes and probed with the indicatedantibodies Proteins were visualized using an ECL detection system

from those that were apoptotic (annexin V-positivePI neg-ative) The percentages of the population reported as viableapoptotic and dead by the Tali cytometer were comparablewith data from the same samples independently run on a flowcytometer

26 Statistical Analysis Statistical significance was deter-mined through analysis of variance (ANOVA)119901 values of lessthan 005 were considered statistically significant

3 Results and Discussion

31 Effects of A asiatica Extract and Eupatilin on Cisplatin-Induced Nephrotoxicity in LLC-PK1 Cells The antioxidanteffects of A asiatica and eupatilin were tested using DPPHa stable free radical DPPH decolorizes in the presenceof antioxidants The scavenging ability of A asiatica andeupatilin was represented by a line diagram and com-pared with vitamin C (Figure 1(b)) This result suggests thateupatilin is the antioxidant and active component of Aasiatica As shown in Figure 1 the cell viability was decreased


Control Cisplatin Cisplatin + Artemisia 250120583gmL

Cisplatin + eupatilin 10 120583gmL Cisplatin + eupatilin 50120583gmL

(a)

12

10

8

6

4

2

0

Control Cisplatin Cisplatin +

Artemisia250120583gmL

Cisplatin +

eupatilin10 120583gmL

Cisplatin +eupatilin50120583gmL

Apop

totic

cells

()

(b)

Figure 3 Effects of A asiatica extract and eupatilin on apoptosis in LLC-PK1 cells (a) Representative images of apoptosis detection(b) Percentage of annexin V-positive-stained apoptotic cells Dead and apoptotic cells were stained red and green respectively Apoptosiswas determined by a Tali image-based cytometer

significantly to about 60 of that of untreated control cells(119901 = 00004) after 25120583M cisplatin treatment However pre-treatment with the A asiatica extract and eupatilin markedlyrestored cell viability to 80 and 82 respectively in a dose-dependent manner (Figures 1(c) and 1(d))

32 Involvement of MAPKs-Caspase-3 Signaling Pathway inthe Protective Effect of A asiatica Extract and Eupatilin againstCytotoxicity in Cultured LLC-PK1 Cells Figure 2 shows theprotein expressions of p38 p-p38 JNK p-JNK ERK p-ERK and cleaved caspase-3 afterA asiatica (250120583gmL) andeupatilin (10 and 50 120583gmL) treatment As shown in Figure 2the phosphorylation of p38 and JNK was decreased in LLC-PK1 cells by A asiatica and eupatilin treatments In addition

the elevated protein expression of cleaved caspase-3 was alsomarkedly reduced by A asiatica and eupatilin treatmentsERK protein expression in the LLC-PK1 cells was slightlydecreased by cisplatin treatment and increased by A asiaticaand eupatilin treatment however the differences were not ofsignificant effect

33 Effects of A asiatica Extract and Eupatilin on Apoptosis inLLC-PK1 Cells Figure 3 shows the effects of the A asiaticaextract and eupatilin on apoptosis in LLC-PK1 cells Asshown in Figure 3(a) the number of dead and apoptoticcells which were stained with red or green colors wasincreased by cisplatin treatment whereas it was decreasedafter cotreatment with the A asiatica extract (119901 = 0008)


and more significantly eupatilin (119901 = 0003) The elevatedpercentage of apoptotic cells after cisplatin treatment wasmarkedly decreased after cotreatment with the A asiaticaextract and eupatilin (Figure 3(b))

4 Discussion

The protective effect of the A asiatica extract and eupatilinagainst cisplatin-induced nephrotoxicity was tested usingLLC-PK1 cells which are the most vulnerable renal tubularcells to oxidative stress [24 28] It is reported that reactiveoxygen species (ROS) play vital biological roles in cellularhomeostasis whereas the increased ROS levels are associatedwith apoptosis in cells [29ndash31] Our result suggests thateupatilin is the antioxidant and active component of Aasiatica In addition pretreatment with theA asiatica extractand eupatilin markedly ameliorated reduced LLC-PK1 cellviability by cisplatin in a dose-dependent manner

It has been reported that ROS act as a second messengerinitiating signal transduction cascades including theMAPKssignaling pathway [32 33] The MAPKs are important medi-ators for apoptosis induction in response to anticancer drugsin particular cisplatin [34 35] In the present study theincreased protein expressions of phosphorylated JNK andp38 by cisplatin in LLC-PK1 cells were markedly amelioratedafter A asiatica extract or eupatilin cotreatment To furtherinvestigate the ability of A asiatica and eupatilin to preventapoptosis we measured the expression levels of cleavedcaspase-3 known as an index of apoptosis in the kidney Asshown in the results A asiatica and eupatilin significantlyreduced the expression of cleaved caspase-3 These resultssuggest that cisplatin-induced increases in the ROS levelactivate MAPKs in LLC-PK1 cells whereas the A asiaticaextract and eupatilin inhibit the activation of p38 and JNK

Cisplatin-induced renal cell damage is dependent onapoptosis induced by DNA damage [36 37] Apoptosis inrenal tubular cells has been observed in several kinds ofrenal disorders [38] The elevated protein level of cleavedcaspase-3 decreased after treatment with A asiatica extractand eupatilin

In conclusion our results suggest that the A asiaticaextract can ameliorate nephrotoxicity in LLC-PK1 cellsEupatilin serves as one of the major components by blockingthe MAPKs-caspase-3 signaling cascade

Conflict of Interests

The authors declare no conflict of interests in this work

Authorsrsquo Contribution

Jun Yeon Park and Dahae Lee contributed equally to thecontent of this paper

Acknowledgments

This research was funded by the Gangneung Asan HospitalBiomedical Research Center Promotion FundThis work was

also supported by a grant from Korea Institute of Science andTechnology Republic of Korea (2Z04371)

References

[1] M H Hanigan and P Devarajan ldquoCisplatin nephrotoxicitymolecular mechanismsrdquoCancerTherapy vol 1 pp 47ndash61 2003

[2] J Li K Jiang X Qiu et al ldquoOverexpression of CXCR4is significantly associated with cisplatin-based chemotherapyresistance and can be a prognostic factor in epithelial ovariancancerrdquo BMB Reports vol 47 no 1 pp 33ndash38 2014

[3] M E I Leibbrandt G H IWolfgang A LMetz A A Ozobiaand J R Haskins ldquoCritical subcellular targets of cisplatin andrelated platinum analogs in rat renal proximal tubule cellsrdquoKidney International vol 48 no 3 pp 761ndash770 1995

[4] W Lieberthal V Triaca and J Levine ldquoMechanisms of deathinduced by cisplatin in proximal tubular epithelial cells apop-tosis vs necrosisrdquoThe American Journal of Physiology vol 270no 4 pp F700ndashF708 1996

[5] A H Lau ldquoApoptosis induced by cisplatin nephrotoxic injuryrdquoKidney International vol 56 no 4 pp 1295ndash1298 1999

[6] H R Brady B C Kone M E Stromski M L Zeidel GGiebisch and S R Gullans ldquoMitochondrial injury an earlyevent in cisplatin toxicity to renal proximal tubulesrdquo TheAmerican Journal of PhysiologymdashRenal Fluid and ElectrolytePhysiology vol 258 no 5 pp F1181ndashF1187 1990

[7] H Huang L Zhu B R Reid G P Drobny and P B HopkinsldquoSolution structure of a cisplatin-induced DNA interstrandcross-linkrdquo Science vol 270 no 5243 pp 1842ndash1845 1995

[8] H Jiang X Liao and P Li ldquoExperimental study on apoptosisand Apo-1 expression of buccal carcinoma cell (BCC) inducedby cisplatinrdquoHuaxiKouqiangYixue Zazhi vol 17 no 4 pp 300ndash303 1999

[9] H Zhou T Miyaji A Kato Y Fujigaki K Sano and AHishida ldquoAttenuation of cisplatin-induced acute renal failure isassociated with less apoptotic cell deathrdquo Journal of Laboratoryand Clinical Medicine vol 134 no 6 pp 649ndash658 1999

[10] F Shiraishi L M Curtis L Truong et al ldquoHeme oxygenase-1gene ablation or expression modulates cisplatin-induced renaltubular apoptosisrdquo The American Journal of PhysiologymdashRenalPhysiology vol 278 no 5 pp F726ndashF736 2000

[11] H N Varzi S Esmailzadeh H Morovvati R Avizeh AShahriari and M E Givi ldquoEffect of silymarin and vitaminE on gentamicin-induced nephrotoxicity in dogsrdquo Journal ofVeterinary Pharmacology and Therapeutics vol 30 no 5 pp477ndash481 2007

[12] G Harlalka C Patil and M Patil ldquoProtective effect ofKalanchoe pinnata pers (Crassulaceae) on gentamicin-inducednephrotoxicity in ratsrdquo Indian Journal of Pharmacology vol 39no 4 pp 201ndash205 2007

[13] J C Jeong W M Hwang C H Yoon and Y K Kim ldquoSalviaeradix extract prevents cisplatin-induced acute renal failure inrabbitsrdquo Nephron vol 88 no 3 pp 241ndash246 2001

[14] T Y Oh G J Ahn S M Choi B O Ahn and W B KimldquoIncreased susceptibility of ethanol-treated gastric mucosa tonaproxen and its inhibition by DA-9601 an Artemisia asiaticaextractrdquo World Journal of Gastroenterology vol 11 no 47 pp7450ndash7456 2005

[15] B K Ryu B O Ahn T Y Oh S H Kim W B Kim and E BLee ldquoStudies on protective effect of DA-9601 artemisia asiaticaextract on acetaminophen- and CCl

4

-induced liver damage in


ratsrdquo Archives of Pharmacal Research vol 21 no 5 pp 508ndash5131998

[16] K-B Hahm J-H Kim B-M You et al ldquoInduction of apoptosiswith an extract of Artemisia asiatica attenuates the severity ofcerulein-induced pancreatitis in ratsrdquo Pancreas vol 17 no 2 pp153ndash157 1998

[17] H J Seo K K Park S S Han et al ldquoInhibitory effects of thestandardized extract (DA-9601) of Artemisia asiatica Nakai onphorbol ester-induced ornithine decarboxylase activity papil-loma formation cyclooxygenase-2 expression inducible nitricoxide synthase expression and nuclear transcription factor 120581Bactivation in mouse skinrdquo International Journal of Cancer vol100 no 4 pp 456ndash462 2002

[18] T Y Oh J S Lee B O Ahn et al ldquoOxidative stress is moreimportant than acid in the pathogenesis of reflux oesophagitisin ratsrdquo Gut vol 49 no 3 pp 364ndash371 2001

[19] S Y Seol M H Kim J S Rew and M G Choi ldquoA phase IIIclinical trial of Stillen(TM) for erosive gastritisrdquo Korean Journalof Gastrointestinal Endoscopy vol 28 no 5 pp 230ndash236 2004

[20] E B Lee S A Cheon E S Lee et al ldquoGeneral pharmacologyof artemisia extract powder DA-9601rdquo Journal of AppliedPharmacology vol 4 no 2 pp 174ndash183 1996

[21] T Y Oh B O Ahn J I Ko et al ldquoStudies on protective effect ofda-9601 an artemisiae herba extract against ethanol-inducedgastric mucosal damage and its mechanismrdquo Biomolecules ampTherapeutics vol 5 no 2 pp 202ndash210 1997

[22] E-J Choi H-M Oh H Wee et al ldquoEupatilin exhibits anovel anti-tumor activity through the induction of cell cyclearrest and differentiation of gastric carcinoma AGS cellsrdquoDifferentiation vol 77 no 4 pp 412ndash423 2009

[23] J-H Cheong S Y Hong Y Zheng and S H Noh ldquoEupatilininhibits gastric cancer cell growth by blocking STAT3-mediatedVEGF expressionrdquo Journal of Gastric Cancer vol 11 no 1 pp16ndash22 2011

[24] T Yokozawa E J Cho Y Hara and K Kitani ldquoAntioxidativeactivity of green tea treated with radical initiator 221015840-azobis(2-amidinopropane) dihydrochloriderdquo Journal of Agricultural andFood Chemistry vol 48 no 10 pp 5068ndash5073 2000

[25] H-Y Pan Y Qu J-K Zhang T G Kang and D-Q DouldquoAntioxidant activity of ginseng cultivated under mountainousforest with different growing yearsrdquo Journal of Ginseng Researchvol 37 no 3 pp 355ndash360 2013

[26] K I Song J Y Park S Lee et al ldquoProtective effect oftetrahydrocurcumin against cisplatin-induced renal damage invitro and in vivo studiesrdquo Planta Medica vol 81 no 4 pp 286ndash291 2015

[27] A Dubey J W Min H J Koo et al ldquoAnticancer potency andmultidrug-resistant studies of self-assembled arene-rutheniummetallarectanglesrdquo ChemistrymdashA European Journal vol 19 no35 pp 11622ndash11628 2013

[28] K S Kang J Ham Y-J Kim J H Park E-J Cho and NYamabe ldquoHeat-processed Panax ginseng and diabetic renaldamage active components and action mechanismrdquo Journal ofGinseng Research vol 37 no 4 pp 379ndash388 2013

[29] I Dolado A Swat N Ajenjo G De Vita A Cuadrado and AR Nebreda ldquop38120572 MAP kinase as a sensor of reactive oxygenspecies in tumorigenesisrdquo Cancer Cell vol 11 no 2 pp 191ndash2052007

[30] J Wang and J Yi ldquoCancer cell killing via ROS to increase ordecrease that is the questionrdquo Cancer Biology ampTherapy vol 7no 12 pp 1875ndash1884 2008

[31] S Zhang Y Sun Z Yuan et al ldquoHeat shock protein 90120573 inhibitsapoptosis of intestinal epithelial cells induced by hypoxiathrough stabilizing phosphorylated Aktrdquo BMB Reports vol 46no 1 pp 47ndash52 2013

[32] S Ramachandiran Q Huang J Dong S S Lau and TJ Monks ldquoMitogen-activated protein kinases contribute toreactive oxygen species-induced cell death in renal proximaltubule epithelial cellsrdquo Chemical Research in Toxicology vol 15no 12 pp 1635ndash1642 2002

[33] H-S Lee G-S Lee S-H Kim H-K Kim D-H Suk andD-SLee ldquoAnti-oxidizing effect of the dichloromethane and hexanefractions from Orostachys japonicus in LPS-stimulated RAW2647 cells via upregulation of Nrf2 expression and activationof MAPK signaling pathwayrdquo BMB Reports vol 47 no 2 pp98ndash103 2014

[34] I H Bae S W Kang S H Yoon and H-D Um ldquoCellularcomponents involved in the cell death induced by cisplatin inthe absence of p53 activationrdquo Oncology Reports vol 15 no 5pp 1175ndash1180 2006

[35] I Sanchez-Perez M Martınez-Gomariz D Williams S MKeyse and R Perona ldquoCL100MKP-1modulates JNK activationand apoptosis in response to cisplatinrdquoOncogene vol 19 no 45pp 5142ndash5152 2000

[36] J W Pippin R Durvasula A Petermann K Hiromura W GCouser and S J Shankland ldquoDNA damage is a novel responseto sublytic complement C5b-9-induced injury in podocytesrdquoThe Journal of Clinical Investigation vol 111 no 6 pp 877ndash8852003

[37] K S Kang W Lee Y Jung et al ldquoProtective effect of esculin onstreptozotocin-induced diabetic renal damage in micerdquo Journalof Agricultural and Food Chemistry vol 62 no 9 pp 2069ndash2076 2014

[38] N Ueda G P Kaushal and S V Shah ldquoApoptotic mechanismsin acute renal failurerdquo American Journal of Medicine vol 108no 5 pp 403ndash415 2000

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


HO

H3CO

OOH

OCH3

OCH3

O

(a)

0

20

40

60

80

100

120

0 10 25 50 100

DPP

H ra

dica

l sca

veng

ing

activ

ity

Vitamin CArtemisiaEupatilin

Concentration (120583gmL)

(b)

120

100

80

60

40

20

0

0

10

25

50

100

250


Con

trol

Cel

l via

bilit

y (

of c

ontro

l)

Artemisia (120583gmL)

(c)

120

100

80

60

40

20

0

0

10

25

50

100


Con

trol

Cel

l via

bilit

y (

of c

ontro

l)

Eupatilin (120583gmL)

(d)

Figure 1 Effects of A asiatica extract and eupatilin on cisplatin-induced nephrotoxicity in LLC-PK1 cells (a) Structure of eupatilin(b) Comparison of DPPH radical scavenging effects ofA asiatica extract eupatilin and vitamin C (c) Dose-dependent protective effect ofAasiatica extract against cisplatin-induced nephrotoxicity in cells (d) Dose-dependent protective effect of eupatilin against cisplatin-inducednephrotoxicity in cells

Stillen is a commercially available extract fromA asiaticaEupatilin (Figure 1(a)) an active compound isolated fromA asiatica has been reported to treat peptic ulcers andgastritis It has antioxidative and anti-inflammatory effects

against gastric mucosal injury [18 19] Various inflammatorymediators such as cytokines and oxidative stress that canaffect gastric mucosal injury are thought to be involved inits action mechanism [20 21] Eupatilin was also reported to








0minus

119860

1)119860




of the extract



minus

minusminus

+

minusminus

+

minus+

+

+minus


+

minusminus

+

Cleaved caspase-3

p-p38

GAPDH

p38

p-ERK

ERK

p-JNK

JNK











(a)

12

10

8

6

4

2

0



Cisplatin +



Apop

totic

cells

()

(b)








4 Discussion









Acknowledgments



References
















4
































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of























0minus

119860

1)119860




of the extract



minus

minusminus

+

minusminus

+

minus+

+

+minus


+

minusminus

+

Cleaved caspase-3

p-p38

GAPDH

p38

p-ERK

ERK

p-JNK

JNK











(a)

12

10

8

6

4

2

0



Cisplatin +



Apop

totic

cells

()

(b)








4 Discussion









Acknowledgments



References
















4
































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















(a)

12

10

8

6

4

2

0



Cisplatin +



Apop

totic

cells

()

(b)








4 Discussion









Acknowledgments



References
















4
































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















4 Discussion









Acknowledgments



References
















4
































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of














































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















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