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Original Article ฤทธติ์ า้ นอนุมูลอิสระ และความเป็นพษิ ต่อเซลล์ของสารสกัดจากพชื Camellia nitidissima Chi. Hua Zhu1, Xu Zhao1, บงั อร ศรพี านชิ กุลชัย2*, แคทรียา สทุ ธานุช2, Xiao-Xun Wang1 บทคดั ย่อ ฤทธติ์ ้านอนุมลู อสิ ระ และความเป็นพษิ ต่อเซลล์ของสารสกดั จากพืช Camellia nitidissima Chi. Hua Zhu, Xu Zhao, บงั อร ศรพี านชิ กลุ ชยั , แคทรียา สุทธานชุ , Xiao-Xun Wang ว. เภสัชศาสตร์อีสาน 2554; 7(1) : 11-17 Received: 17 March 2010 Accepted : 2 March 2011 บทนำ�: Camellia nitidissima Chi. เป็นพืชสมุนไพรของจีน เรียกว่า “จินฮัวชา” ในยาแผนโบราณของจีน โดย ใช้ส่วนใบในการรักษาโรคตับ, ฝีอักเสบ, ท้องเสีย, ความดันโลหิตสูง, และเนื้องอก ในการศึกษาครั้งนี้มีวัตถุประสงค์เพื่อ ตรวจสอบฤทธ์ทิ างชวี ภาพ ฤทธิ์ต้านอนุมลู อิสระ และความเป็นพษิ ต่อเซลลข์ องสารสกัดจากใบ Camellia nitidissima Chi. วสั ดุและวธิ กี ารทดลอง: โดยเตรยี มส่วนสกัดชนั้ ต่างๆ จากผงแห้งของใบ Camellia nitidissima Chi. โดยวิธกี ารหมักผง ใบแหง้ ในตัวท�ำ ละลายชนิดตา่ งๆ ได้แก่ 70% เอทานอล (F1), 40 – 60% อเี ทอร์ (F2), เอทิลอะซิเตต (F3), เอน็ -บวิ ทานอล (F4) และนํา้ (F5) แลว้ ตรวจสอบฤทธิต์ ้านอนมุ ลู อสิ ระ โดยใช้ DPPH assay และความเปน็ พิษตอ่ เซลล์ โดยใช้ MTT assay ผลการศกึ ษา: ผลการศกึ ษาพบวา่ ความสามารถในการออกฤทธขิ์ องสว่ นสกดั ชน้ั ตา่ งๆ มคี วามแตกตา่ งกนั โดยสว่ นสกดั ชน้ั นา้ํ (F5) แสดงฤทธต์ิ า้ นมนมุ ลู อสิ ระทด่ี กี วา่ สว่ นสกดั ชนั้ อนื่ ๆ (EC50 = 63.31 ppm) ในขณะสว่ นสกดั ชน้ั เอน็ -บวิ ทานอล (F4) แสดงความเปน็ พษิ ตอ่ เซลล์ ดว้ ยคา่ IC50 เทา่ กบั 184.0 µg/ml ซงึ่ เปน็ สว่ นสกดั ทค่ี วรทำ�การศกึ ษาเพมิ่ เตมิ เกย่ี วกบั ฤทธติ์ า้ น เซลลม์ ะเร็งตอ่ ไป สรุปผล: การศึกษานพี้ บว่า Camellia nitidissima Chi. แสดงฤทธติ์ ้านการเกิดออกซิเดชันและตา้ นเซลล์ มะเรง็ ตับ HepG2 ซึง่ ควรมกี ารศกึ ษาเพมิ่ เตมิ เก่ยี วกบั กลไกการออกฤทธิ์, ประสิทธิภาพในการรกั ษา, และความปลอดภัย ค�ำ ส�ำ คัญ: Camellia nitidissima Chi., ฤทธ์ิต้านอนมุ ูลอสิ ระ, ฤทธคิ์ วามเปน็ พิษตอ่ เซลล,์ DPPH assay, MTT assay Abstract Antioxidative and Cytotoxic Effect of the Extract from Camellia nitidissima Chi. Hua Zhua, Xu Zhaoa, Bungorn Sripanidkulchaib,*, Khaetthareeya Sutthanutb, Xiao-Xun Wanga IJPS, 2011; 7(1) : 11-17 Introduction: Camellia nitidissima Chi. leaves or yellow camellia is a traditional Chinese herb called “Jin Hua Cha” in Chinese medicine. It is commonly used for the treatment of liver diseases, faucitis, diarrhea, edema, hypertension, and tumor. This study aimed to investigate the antioxidative and cytotoxic effects of the extracts from 1 Faculty of Pharmacy, Guangxi Traditional Chinese Medical University, Nanning, China. 2 ศูนย์วิจยั และพัฒนาผลติ ภัณฑส์ ุขภาพจากสมุนไพร คณะเภสชั ศาสตร์ มหาวทิ ยาลัยขอนแก่น * ติดต่อผนู้ ิพนธ:์ โทรศพั ท/์ โทรสาร 043-203149 e-mail: [email protected] 2 Center for Research and Development of Herbal Health Products. Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand. * Corresponding author: Tel/Fax 043-203149 e-mail: [email protected]

Antioxidative and Cytotoxic Effect of the Extract from Camellia nitidissima Chi. IJPS 12 Zhu H. et al. Vol. 7, No1, Jan - April 2011 Camellia nitidissima Chi. leaves. Materials and Method: Dried Camellia nitidissima Chi. leaves were extracted with 70% ethanol (F1), then further extracted with 40-60% petroleum ether (F2), ethyl acetate (F3), n-Butanol (F4) and water (F5) individually to obtain 5 fractions. The plant extracts were determined for antioxidative activity by DPPH assay, and cytotoxicity effect to HepG2 hepatoma cell line by MTT assay. Results: The results demonstrated the different potency of action among the different extracts. The F5 (water fraction) had notable anti-oxidative potential wfuirtthheErCs5t0udoife6d3f.o3r1apnpti-mca. nBcaesreadctoivnityth.eColonwcelustsIiCon50: at 184.0 µg/ml of F4 (n -butanol fraction) this fraction should be The antioxidant properties and cytotoxicity to a HepG2 hepatoma cell line of Camellia nitidissima Chi. leave extracts were studied. However, further studies should consider identifying related mechanisms, efficacy of action, and safety for use. Keywords: Camellia nitidissima Chi., Antioxidative effect, Cytotoxic effect, DPPH assay, MTT assa Introduction China (Fu, 1992). To support the conservation and management programs for C. nitidissima, currently, a Camellia nitidissima Chi., the yellow camellia yellow Camellia public park was set up in Nanning of belonging to Theaceae family, is one of rare plants first Guangxi and this park becomes a new sight spot in discovered in 1933 (Deng et al., 2000). It is distributed in Nanning (Yan et al., 2003). a narrow region of Guangxi Province, South China, and In traditional Chinese medicine, C. nitidissima North Vietnam, where it grows in acidic soils along Chi. leaves have the effects of heat-clearing and shady and moist valley under evergreen forests at the detoxication, strangury-relieving, anti-cancer, inhibition latitudes of 50–650 m (Wei et al., 2008; Yang et al., of tumor growing, reducing blood lipid, preventing 2008). In the early 1970s, C. nitidissima was honored as atherosclerosis, increasing the immune function, and “the queen of camellias family” (Liang, 1993); It is one of draining dampness (Fu et al., 2005). Normally, it uses the typical species in sect nitidissima (Ye and Xu, 1992). to treat faucitis, diarrhea, edema, hypertension, and C. nitidissima has the big entomogamous flower, golden, prevent liver disease and tumor (Huang, 1999). C. and transparent waxy petal (Liang, 1993). It has been nitidissima Chi. leaves contain several of trace elements: introduced to Japan, Australia, and North America as a germanium, selenium, manganese, molybdenum, vanadium, useful genetic resource for commercial cultivation and zinc, polyphenols, and essential amino acid. ofcamellias, attracting extensive attention of horticultural In the recent years, the extract from the plant workers worldwide (Nishimoto et al., 2004; Parks, 2000). belonging to same genus, Camellia chrysantha was In addition, it has been reported that the leaves, flowers, reported to have remarkable antioxidation properties to and seed oils of C. nitidissima can be of value in food the free radical which is produced by the Fenton reacting and Chinese traditional medicine (Liang, 1993). In system and pyrogallic acid autoxidation (Qin et al., 2008; China, natural populations of camellias including C. Yan and Yao, 2009). Moreover, the leaves of C. chrysantha nitidissima have been investigated extensively for show inhibitory effect on the DEN-induced precancerous several decades. C. nitidissima has been found only in lesion of rat liver with dose-dependent manner (Duan two disjunctive areas of Guangxi: one is in the junction of et al., 2006) and the human hepatoma cells BEL-7404 Fushu, Longan, and Fusui near Nanning City, and the cultured in vitro (Li et al., 2007). Therefore, this study other is in Fangcheng, south of Mount Shiwan (Su and aims to determine antioxidative and cytotoxic activity Mou, 1988). Due to deforestation and collection of seeds to the human hepatoma cell line (HepG2 cells) of for horticulture, its natural population has declined C. nitidissima crude extracts by using DPPH and MTT dramatically in recent decades. C. nitidissima has been assays, respectively. included in the checklist of State Protection Category I in

ว. เภสัชศาสตร์อสี าน ฤทธิ์ตา้ นอนมุ ลู อสิ ระ และความเปน็ พิษต่อเซลล์ของสารสกดั จากพชื Camellia nitidissima Chi. 13 ปีที่ 7 ฉบบั ท่ี 1 ม.ค. - เม.ย. 2554 Zhu H. et al. Methods of 70% ethanol for 48 h, and twice repeated. Then, the supernatant was collected and filtered through Whatman 1. Chemicals no.1 filter to obtain the filtrate for drying by using rotary 95% ethanol, petroleum ether (Analytical evaporator and freeze dryer. The dried crude extract reagent A.R.), ethyl acetate (HPLC grade) and 1-Butanol was delivered as ‘F1’. The further fractionation was (AR grade) were purchased from Labscan Asia Co.,Ltd. performed by dissolving 20 g of F1 in 350 ml of (Bangkok, Thailand). Dulbecco’s Modified Eagle Medium distilled water and successively partitioned with 350 ml of (DMEM) powder was obtained from Invitrogen corporation. different solvents in order of petroleum ether, ethyl All the other chemicals used were of analytical grade. acetate, and n-Butanol. With % yield of 0.195-5.92, 2. Preparation of herbal extract the dry extracts from each solvent was obtained as C. nitidissima Chi. leaves were collected from AHnutaioZxFdihdrauiaen,tdgievtleCeaaahl.fnepdnogCwyidntoeNtroawxincansEincffgoe,ncGttinouufaotnhuegsxlEyi,xmtCraahccinteafrra.otm5e0dC0iangm6oelfliltiteahrnesitidiss(FimT5aa(baCleqhui1.e)o. us fraction) abd F2, F3 and F4, respectively 12 | Vol. 7 No. 1 Jan - April IJPS 2011 TabTlaeb1leP1hPyshicyaslicaaplpaepapraenacreanacned a%ndyie%ldyoifelCd. onfitiCdi.snsimtidaislesaimveasleaxtvreacstsextracts Fraction Type of solvent for extraction Physical appearance % yield F1 70% Ethanol Rough and dark brown powder 5.92 F2 Petroleum ether Rough and brown powder 0.20 F3 Ethyl acetate 0.72 F4 Viscous and sticky dark purple mass 2.45 F5 n-butanol Rough and yellow powder 1.89 Distilled water Rough and dark yellow powder sf3pIwvrch.eielaDeelkvneocerotrahneilcd2iagcr,i,nmic2nga2-gi( rau3cDel0ncMa.os.0oiadWelpsDiofa6mltidrathcierabo)horaep.ttbdiomeoelnlnoilofIntreysustyvmoscclnviD-oc2fhfrai1idrelno,Pa(aevua-s2eWalepsetnPenni-tnoitdDtiecHoigpnrbrotaciIhinttyepnvaoiowxdesleheghosininicdnkleyirfltpoanoaeaandaclalgribdci.nycrhninlchiaoeulltaticW-i,eznmitcn1oaoy2ygeyhgm-cmx0lpdeee(ati0pWdi,oiln(ncl6vofoDanhtrred)uihonwDy.litPtnygeymttlIPadhPhtaSssc(uPyhHvcMihosdnHacit)oalegoriisfolvlarhaednhrieiibszsttiertdaeeyodyttiieorofoln,aaenivsnn(dpsirodD2gccuasaclo0lcPtudhreSam0eaetiiPgbihcd0nyeomhHaale,)dnen.hlol)fltrinohedilweeiys, cCpdcμaoonelreonnttaiep-fccsoratee4qlzvMmhixrvnnoiu.tieaeihTdinttlDanibrrdeaTsueaangeltademttitrbtoiimifcooeefownyWicennfperbrehemlmlderdlherilsoocoelfieasifhsaffmnnebrn,zsneccaty15aiohdtdetst3h05snleieuv2u0-eos(ilo0rpstnfn4te(iof0aMppinMg,nar5onpmsgμmcgTTd-mafiltDnTiTaoemooCwsrifd)fnmspdypmta1etol,imtesheesdesolmcusettiittthrxhpthrtMoerieteapwyroaoucexdnlunnrpatDtpisdehulrscohsyaPtpic.alcaovrlaPeencoerztiUtctdeoHodnodoastflmamt.mlaoiadi-ivrnon2treiiTmdiensimgnb-tthsesytetyoshaaoedClvrn)oezli.nti-vnlot2la2y,uawe,Tmn,8tmdtet5tiuhhho0hir-iwitseene0ennedoehicpdainhchmuohtemononcdubycarnlehintaetretbcroooaek Currently, 2D0P0P0)H. Cmuerrtehnotdly,hDaPsPbHeemnetuhsoeddhaassboeneen uosfebdaassicone methafonrol.thTehecmyitxottuorxeicwaesfvfeigcotro(uMsloyssshmaakenn, a1n9d8k3e)ptand it was screening of sbtaespicsscrefoerningsestaerpcshifnogr seanrcehwing naenwtioaxnitdioaxnidt ant at roosmuctecmespsefrualtluyreufsoerd15fomrinhueterbsa. Tl heeffeabcst oorbnacnecell (gAr)owth (Liu e compoundscomfrpoomundnsatfurormal nraetsuoraulrcressouirnccelsudinincgludisnpgicsepsi,ces, was male.,as2u0r0e0d).at 515 nm. The results were expressed herbs, fruithse,rabns,dfrvueitgs,eatanbdlevseg(eKtiambleasnd(KKimima,n2d0K0i6m;, L2e0e06e;tLee as EC50 4(5.10%Prienhpiabritaiotniocnonocfehnetrrabtiaolnt)e. st sample al., 2004; eXtual.e, t20a0l.4, ;2X0u05e)t.aTl.,h2e0re0f5o)r.eT,hDerPePfoHre,aDsPsaPyH assay 4. DeterminTathioenpolfaCntytoetxotrxaicctacwtiavsitydissolved with DMSO to described absydSesrcipriabneiddkbuylcShriapianeitdkaul.lch(2a0i 0et8a)l.w(2a0s08u)swedasinused fd tpdhoihirpereomhctmeoatimalnnmziyntcaaoye4drunlckett.hbeee2Wlaowatsrrt.CanhetTheaedeeeTsihccdrdlznheoihltaoeoenachld3ccoitutuuaahe-f3lmm(mnetnil47univtoano,rtbrib5oCuannueern-gtm,oiDhatoq9mb5cenniuo6eem%pidadf-lroalwensetfoMtCtrteoif1hrefT(lOmieelvM0aysTsid20latauTtmbhlffT(mtcbooli1ieen)eyarrgxnmglzlci1/2utssmoeao04sllz(f5llir-sH.nae2ht,gnhed-.cteypuhesCflcGeolp)plees-reu2Mm2ld/lcrm)s,pTte5rwillTdeonw)-,eereraencdturletuathrteeeddn this study.in this study. test waitsh cvoamrmioounsly cuosnecdentotracthioencsk, fowrithetrciyptloictoaxtiiocn for each Follo wing theFomlloewthinogd trheepomretethdodbyreSproiprtaendidbkyulScrhipaai nid concentration, of the herbal extracts ranging between cesbrotyulaudulte.sioin(ne2gsx0t0Drwdskaa8Pouenec)lltd,PrutcesehtHrtismhoat(pneiaiaFnsrnese1eadotspd-nafaaarttbyFhidlr-..yeeo5d(sFx)uc2.oisird0rluwisun0Tadtteg8thlieiyor)oe,,DenensxPttshhtsPraaweeaecHmceptastiraspvaenarils(tmteaiFysp-tao1perseyxw-lleoy.ipFdalaF5uasarpi)ttreniisroowdddetnnle.epysrts,aTeeatrthacrhoesmteneeifdvdipnissattayheabarrmedmdyawtppeallleeys dissolving in methanol to obtain the concentration of 150

Antioxidative and Cytotoxic Effect of the Extract from Camellia nitidissima Chi. IJPS 14 Zhu H. et al. Vol. 7, No1, Jan - April 2011 effect (Mossman, 1983) and it was successfully used for medium was removed. The formazan was dissolved herbal effect on cell growth (Liu et al., 2000). with DMSO and measured the absorbance by using 4.1 Preparation of herbal test sample microplate reader at wavelength 570 nm and 650 nm The plant extract was dissolved with as a reference. The %inhibition and IC50 were calculated DMSO to make a concentration of 100 mg/ml. and expressed as Mean ± SD of IC50. 4.2 Cell culture and treatment Results The human hepatoma cells (HepG2) were cultured and seeded into 96-wells (1x105 cells/ml), and 1. Antioxidant activity then incubated at 37oC, 5% CO2 for 24 h. Cells were As shown in Fig 1, standard vitamin C had treated with various concentrations, with triplication strong antioxidative activity with EC50 of 6.80±0.25 for each concentration, of the herbal extracts ranging ppm (N = 3). Among F1-F5 of C. nitidissima leaf, only between 0-400 μg/ml and incubated for 18 h. Then, the water fraction (F5) was shown to have notable MTT stock solution (5 mg/ml) was diluted to 1 mg/ml in anti-oxidative potential with EC50 of 63.32 ppm (Fig culture medium as a working MTT solution. The treated 2); other fractions showed EC50 greater than 300 ppm cells were added with working MTT solution (50 μl/well) (results not shown). to analyze cell viability. After 3 h incubation, the culture ว. เภสัชศาสตรอีสาน ฤทธ์ติ า นอนุมูลอสิ ระ และความเปนพษิ ตอ เซลลข องสารสกดั จากพืช Camellia nitidissima Chi. | 13 ปท ี่ 7 ฉบบั ท่ี 1 ม.ค - เม.ย. 2554 Hua Zhu, et al. FFiigguurree 11 IInnhhiibbiittiioonn ccuurrvveeooffvviittaammininCCfrfroommDDPPPPHHaasssasyay FigureFi2gu nIrneithi2dibisinItsniiotihimdnibisaictsiuoilmenravacevuleerosavfveferwoosamf ftwreoDarmtPefrPrDafHPcratPiocaHtnsiosan(asFys(5Fa)5y)oof f CC.. 2 cfinc2ifrnre.ae.cacClclClclllusutstyiyiddoottiinowonnnTwssttggehoeo..errxcIxITceenniyiychctttgogeooyaoeapeppbcbtncnillytcsaaseitepavseesirrvilmimraarctvivyllati,,eicmeycldttdhmhoaaeernnopwwddchrchphhohnenhaelaounuorrngcaacloilclcceettgttarreaaeeielcrrrraiaiassstscttelthihheicdccrrasdhiisnnnaokkgowwnfaaefiggtiscgchteeehee,sl,llolttccofahahhhfpeperroHHooopmpeempptptoplaloaaGasGtsntiinn2isi2ntst mwcseotfhrofnaeesdtcrteetecdcpctdantyhoyopesste(tnommaoFHFdttti,oomioeeg5onxxnponwsiisoG3ccs,thartr)a2ee.eatitwfcforneffcFeyeneaddeat4c,octslteltt(msoFtorwoxeFir(gfiiaFcrHm5tnhanie3gb-cdespb)otrf.4imGaufroe)nFt2na.cae4wtlmncldeaoebewoistdllemlrltiestrfhInbbrnC(arobF-fa5bscrtiianngmu0tiscegtoa4ahtanil)nootlw.eonw1ssel8bthnrfdl3eoreIiac.Cwdb9cdyb56ett0eiioonndmptngoaoptottxhmsni1ehcs-,8how3owe.w9er6nde

ว. เภสชั ศาสตรอ์ สี าน ฤทธติ์ ้านอนุมลู อสิ ระ และความเปน็ พษิ ตอ่ เซลล์ของสารสกดั จากพืช Camellia nitidissima Chi. 15 ปีท่ี 7 ฉบบั ท่ี 1 ม.ค. - เม.ย. 2554 Zhu H. et al. Figure 3 Morphological changes of HepG2 cells after treated with 400 µg/ml of C. nitidissima fractions (A = control, B = F1, C = F2, D = F3, E = F4, F = F5).

Antioxidative and Cytotoxic Effect of the Extract from Camellia nitidissima Chi. IJPS IJPS. 14 | 1A6ntioxidaZtihvueHa.nedt Caly. totoxic Effect of the Extract from Camellia nitidissima Chi. Vol. 7, No1, Jan - April 2011 Hua Zhu, et al. Vol. 7 No. 1 Jan - April 2011 DiscussDioisncuasnsdioCnoanncdluCsioonnclusion This stuTdhyisdemstoundsytratdeedmthoenpshtrartmedacotlhoegicaplharmacological potential opfoCte.nntiiatildisosfimCa. lneiatifdeisxstirmacats.leBayf ceoxmtrpaacrtisn.g By comparing EC50 of DEPCP5H0 oafssDayP,PaHn aaqsuseaoyu,safrnacatiqonue(Fo5u)sshfroawcteiodn (F5) showed notable anntiootxaidbaletivaenaticotxividitya,tihvoewaecvteivrityis, theonwtiemveesr lietsis ten times less than standthaardnvsittaamndinaCrd. TvhitiasmfinindinCg. isThciosncfiunrdreinngt wisithconcurrent with the previotuhse reppreovrtiotuhsat rceopmomrtonthlyatfocuonmd manonalqyuefounsd an aqueous extractablexptrhaecntaobliclecompphoeunnodlsicin pclaonmt pleoaufn(dSsripanin- plant leaf idkulchai e(St raipl.,an20id0k8u)l.cUhanideer tthael.c,yto2t0o0xi8c)it.y aUsnsdaeyronthe cytotoxicity hepatomaacseslal lyineon(HheeppGa2to) mbyaMcTeTll alisnseay(,HweepGre2v)eablyedMTT assay, we that n-burteavneoal lefrdacttihoant (Fn4-b) uhtaansosl trofrnagcetisotnac(tFiv4it)y. has strongest Accordingatcotitvhietyir. ICA5c0c,otrhdeinrgelattoivethoredier r IoCf 5c0y,totthoexicriteylative order of was F4 >cFy2tot>oxFic1it>y Fw3a,swhFe4re>asF, 2the>FF51sh>owFe3d, nwohereas, the F5 affect on sHheopwGe2d cneollsa.ffTehcitsomnaHyebpeGa2ttcrieblulste. dThtoistmheay be attributed differencestooftchheemdiicffaelrceonmcepsosiotifoncahmemonicgathl ecforamcptioonssit.ion among the Thereforef,raFc4tiownosu.ldThbeeretfhoerec,aFnd4idwatoeulfdracbteiontheto cbaendidate fraction (o(foimtHfnuroFFrfogoren51tmrtchwea)hehemx-eeadarpitniveFnesilpsdeon4niosortra)tahAtHffoattcmel,ts,ahrorhneloefcaaotsceetanCfeptntbiccbu.wiohstkdvatuc.ttiooerhxaeiieetnltfootatruoineolhvhdnfnnnoamsiTroueeaeotasdliiiwtsrhtrodtnarnprmhhti(,opevhnigo(FheelscufFesecee,rtu5serteaeretnaa1hrdiira)ufulmnfedtntnerhliiaehglsdottsf-itcteaaheifhdlieimamoeececaenonFrballrtcymbslaesdort4utnttssseehditspo)ottcieteeds,niustrwcehnodaonfembehdaiorCexntlonaefstioesausipaiea.utcscntnocuinngsaslfialtnetytnddldietiyptnfdonioafoshottnilchxgtilaotinmhptedirsneeksbinfdoudit.aaesretompouraawtnIsiiohtwnnlftntehiddapiibcoctmeevvdieedtcrteeoeeodhbontoratilnrcrhlliettnvesanofiteisseaarnosciafnvnhanccirflnlaetdofaultcoctogamyeikiiledelccytsnwcnvcimpeamtetoirttgtsoitiioihenohnnlnsnvniasnetdegnteedgscot,.nciaefefxnotshynrriotecuawdattidnnhtfiaytfditehsttogroomeeopisfnbanefoshCettx.tomsweoepw.notanlehoitnttnnieeerIalgtneyrrr.l AcknowfKPTolhraeroeeddnguaRucUeettnshms,ioevreaesFrnrcsawthictsoyuu,ltlaTdynhdlaiokieflaDntPoedhvthfaeoarlromnpskaumctpheepeunottsricttiaanoflgff Herbal Health otShfcisiesntcuedsy,. Khon KPCareoendnteurcUtfnso,irvRDReFeeraesnfcsigetueyrla,GteyyrTceYnhhol,claofaYiewlnPaasdnhncdaDgarefmmoZvraDeecsll,oleuiaLpupmutpiicnoeTarnLtlitcn.SohgAficnHtiehabeni.rsrcibeGesaftsuulr,aHedKnyveg.ihaexolwtinhAofgrriecseBaiorcl hSocni. 2000, 19:119–124. ReferenDcueasn XX, Tang XF, Su JJ. Study on Inhibition of C. Deng GY, Yang cZhDr,ysLaunTthLa. AobnriefDrEevNiewinodfurcetsioenarcohf murine liver on yellocwacnacmere.llJiaMinecdhiRnae.sG, u2a0n0g6x,i3A5g(r6ic):B1io4l-16. FigurFeig4urTeh4e Tinhheibiintihoinbitciounrvecuorfvefivoef frfiavcetiofrnasctoiofnCs. of C. SFcui. 2J0Y0,0,F1u9:1H1J9,–1F2u4. X, et al. Hepu Golden Camellia nitidinsistiidmisasimleavleasvoens HonepHGep2Gc2elclse.lls. Duan XXcc, haTnryacsneagrn.nr2(tXJefhinltoFldaiMadw,tieoisSeodnsIrnun)imsR–DteJhIeEanrJisnp,N.v,ae2Sf2tiso0int0otur0di0ngd5ue6aay.cf,ltftiio3iocon5nan(sc6Iooyny)mfn:hoi1mbpcf4iohut-tiser1oeiin6mnaue.miocplfaoivllCeysor.ptnrhuecntuoCrlsae.maGmcuteiivllilinitay: (A =(FA1=, BF1=, BF2=, FC2,=CF=3,FD3,=DF=4,FE4, =EF=5F).5).

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