다시마추출물의간보호효과 25. Trihloromethyl rdil Trihloromethyl peroxy rdil (CCl 3 OO-), polyenoi ftty id methyl ron (Rekngel, 1976). DNA (Fridovih, 1978). (Hn et

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1 한수지 47(3), 24-21, 214 Originl Artile Kor J Fish Aqut Si 47(3),24-21,214 사염화탄소로유도된흰쥐의간손상으로부터다시마 (Shrin jponi) 추출물의보호효과 김미경 김인혜 남택정 * 부경대학교식품영양학과 Shrin jponi Extrt Protets ginst Cron Tetrhloride-indued Heptotoxiity in Rts Mi-Kyung Kim, In-Hye Kim nd Tek-Jeong Nm* Deprtment of Food Siene nd Nutrition, Pukyong Ntionl University, Busn , Kore The rown lg Shrin jponi is onsumed s foodstuff in mny ountries. Cron tetrhloride ( ) is potent heptotoxin tht is used to ssess heptotoxiity in niml models. This study ssessed the protetive effet of S. jponi extrt (SJE) on -indued ute liver injury in rts. Experimentl rts were divided into the following three groups: ontrol,, nd +SJE; the ltter two groups were given 15 or 3 mg SJE/kg orlly for 1 dys. Three hours fter the finl tretment, ll rts-exept for those in the ontrol group-were dministered intrperitonel injetions of. One dy lter, lood nd liver smples were olleted for evlution iomrker of levels. Asprtte trnsminse (APT; GPT) nd lnine minotrnsferse (ALT; GOT) levels were mrkedly lower in the +SJE group thn the group. The hepti superoxide dismutse (SOD) tivity of the +SJE group ws signifintly lower thn tht of the group. In omprison, glutthione S-trnsferse (GSH) nd tlse (CAT) levels were signifintly higher in the +SJE group. Western lotting reveled tht SJE ttenuted the -indued EGFR nd MAPK tivity in the liver. Thus, we onlude tht SJE protets ginst -indued heptotoxiity. Key words : Shrin jponi, Cron Tetrhloride, GOT/GPT, GSH 서론 (Shrin jponi: se tngle),,,,,., (7-8%), fuoidn lminrin (Hroun-Bouhedj et l., 2; Lee et l., 1998).,,,, (Nishino et l., 1991; So et l., 27; Hng et l,. 25).,, (Gru et l., 2). (Cron tetrhloride, ), (Roins, 1987). mirosome NADPH-ytohrome P-45 eletron trnsport hin ron-hlorine trihloromethyl rdil (CCl 3 ) hlorine rdil (Cl - ) Kor J Fish Aqut Si 47(3) 24-21, June 214 This is n Open Aess rtile distriuted under the terms of the Cretive Commons Attriution Non-Commeril Liens ( permits unrestrited non-ommeril use, distriution, nd reprodution in ny medium, provided the originl work is properly ited. Reeived 16 April 214; Revised 25 April 214; Aepted 3 April 214 *Corresponding uthor: Tel: Fx: E-mil ddress: nmtj@pknu..kr Copyright 214 The Koren Soiety of Fisheries nd Aquti Siene 24 pissn: , eissn:

2 다시마추출물의간보호효과 25. Trihloromethyl rdil Trihloromethyl peroxy rdil (CCl 3 OO-), polyenoi ftty id methyl ron (Rekngel, 1976). DNA (Fridovih, 1978). (Hn et l., 1995; Prk et l., 1991).. 실험재료 재료및방법,,., (S. jponi Extrt, SJE) 4 g 1 L 3. (Supr-22K, Hnil, Kore) 4,416 g, 4, % ethyl lohol 4, overnight Whtmn filter pper (No. 3). (EYELA N-N series, Tokyo rikkiki o., LTD, JAPAN), (EYELA FDU-21, Tokyo rikkiki o., LTD, JAPAN). 실험동물의사육 15 1 g 5 Sprue Dwley rt (SAMTAKO Co., LTD, Gyeonggio-do, Kore), 1 ( 2 2, 5 5%, 12 ) stinless ge () ( ) mouse, ml/rt. 15, 3 mg/kg B.W. 1. 3, 1 ml/kg B.W ethyl ether (2,5 g, 4, 2 min). -7. 혈청중의 GOT/GPT 측정 SJE GOT/GPT Reitmn-Frnkel kit (ASAN PHARM. Co., LTD., Seoul, Kore). GOT/GPT L, 37 GOT 6, GPT 3. 1 L 2.4 N NOH 1 ml 1, ELISA reder (Benhmker, Bio-Rd, Rihmond, CA, USA) 55 nm. 간조직의 Glutthion (GSH) 함량측정 SJE GSH glutthione ssy kit (Sigm- Aldrih Co. LLC., MO, USA)..5 g 5% 5-sulfosliyli id homogenizer , g 15 GSH. working mixture (1 mm potssium phosphte uffer, ph 7., 1 mm EDTA, 6 untis/ml glutthione redutse, 1.5 mg/ml DTNB stok solution) 5.16 mg/ml NADPH 5 ELISA plte reder 412 nm 1. 간조직의 Superoxide dismutse (SOD) 활성측정 SJE SOD superoxide dismutse ssy kit (Cymn Chemil Co., MI, USA)..5 g extrtion uffer (2 mm HEPES, ph 7.2, 1 mm EGTA, 21 mm mnnitol, 7 mm surose) homogenizer , g 15 SOD. rdil detetor solution (5 mm Tris-HC1, ph 8.,.1 mm DTPA,.1 mm hypoxnthine,.25 mg/ml tetrzolium slt solution) 5.25 mg/ ml xnthine oxidse 2 ELISA plte

3 26 김미경ㆍ김인혜ㆍ남택정 reder 45 nm. SOD U/ ml. 간조직의 Ctlse (CAT) 활성측정 SJE CAT tlse ssy kit (Cymn Chemil Co., MI, USA)..5 g extrtion uffer (5 mm potssium phosphte, ph 7., 1 mm EDTA) , g 15 CAT. ssy uffer (1 mm potssium phosphte, ph 7., 3% methnol) 5.35 M/mL hydrogen peroxide 2. 1 M potssium hydroxide.5 M potssium periodte ELISA plte reder 54 nm. Western lot 에의한단백질발현분석 SJE..1 mg RIPA extrtion uffer (5 mm Tris- HCl, ph 7.4, 15 mm NCl, 1% NP-4, 1 mm EDTA, 1 mm NF, 1 mm N 3 VO 4, 1 g/ml protinin, 1 g/ml leupeptin, 1 g/ml pepsttin, 1 mm PMSF,.25% sodium deoxyholte) homogenizer. (12, g, 4, 2 min), sodium dodeyl sulfte-polyrylmide gel eletrophoresis (SDS-PAGE) polyvinylidene fluroide (PVDF) memrne (Milipore, USA). Memrne 1% BSA Tris-uffered sline ontining Tween 2 (TBS-T; 1 mm Tris-HCl, 15 mm NCl,.1% Tween 2, ph 7.5) loking 1 TBS-T 1:1, 2., 2 TBS-T 1:1, 2 super signl west pio stle peroxide solution super signl west pio luminol/enhner solution (Rokford, IL, USA) KODAK X-ry film densitometer (FujiFilm, JA- PAN). 통계처리 (men S.D.), SPSS 1. (Sttistil Pkge for Soil Siene, SPSS In., IL, USA) Onewy-ANOVA test, P<.5 Dunn's multiple rnge test. 결과및고찰 혈청중의 GOT 및 GPT 활성변화 GOT GPT. GOT,, 3. GPT,, 3 GPT (Fig. 1). Cron tetrhloride, thioetmide, dimethyl nitrosmine llyl lohol minotrnsferse (Blze et l., 1961; Reitmn A B GOT (IU/L) GPT (IU/L) Experimentl group Fig. 1. Effets of SJE on serum GOT nd GPT tivities in - indued rt. is ontrol group without dministrtion. (distilled wter 1.5 ml/rt) nd, 3 (S. jponi Extrt 15, 3 mg/kg rt, i.g) group were orl dministrted to rt for 1 dys efore tretment. Different supersripts represent signifintly different results (P<.5), Experimentl group

4 다시마추출물의간보호효과 27 nd Flnkel, 1957). (ULF), GOT GPT 3.3, 3.7, ULF 29.8%, 48.2% (Nm et l., 27). GOT GPT,., 15 mg/ kg, 3 mg/kg GOT, GPT. 간조직중의 GSH 함량변화 GSH.,,, 3 GSH (Fig. 2). GSH Glutthione peroxidse (GPX) Glutthione S- trnsferse (GST) (Hyes et l., 2). GSH GPX glutthione disulfide (GSSG) GSH. GSSG (Vendemile et l. 1989). GSH (Hidk et l., 27), GSH. Chlorell vulgris (CVE) (Kim, 29)., GSH, GSH., 3 GSH SJE GSH, GSH hydrogen peroxide (H 2 ). 간조직중의 SOD 활성변화 superoxide rdil SOD.,,, 3 SOD (Fig. 3). SOD mitohondri superoxide rdil hydrogen peroxide (H 2 ) (Crpo et l., 1978). SOD (Jung et l., 24),. shizndrin C iphenyldimethyl diroxylte (DDB) DWP-4 (DDB : selenium yest : glutthione = 31.1 : GSH level (%) SOD tivity (U/mL) Experimentl group Fig. 2. Effets of SJE on hepti GSH ontent in -indued rt. is ontrol group without dministrtion. (distilled wter 1.5 ml/rt) nd, 3 (S. jponi Extrt 15, 3 mg/kg rt, i.g) group were orl dministrted to rt for 1 dys efore tretment. Different supersripts represent signifintly different results (P<.5). Experimentl group Fig. 3. Effets of SJE on hepti SOD tivities in -indued rt. is ontrol group without dministrtion. (distilled wter 1.5 ml/rt) nd, 3 (S. jponi Extrt 15, 3 mg/kg rt, i.g) group were orl dministrted to rt for 1 dys efore tretment. Different supersripts represent signifintly different results (P<.5).

5 28 김미경ㆍ김인혜ㆍ남택정 : 62.1, w/w%) SOD, DWP-4 (Prk et l., 25). SOD, SOD (Lee, 21). SOD,., 3 SOD. 간조직중의 CAT 활성변화 CAT.,, 3 (Fig. 4). CAT peroxisome, H 2 (H 2 O) ( ) (Sunde nd Hoekstr, 198). CAT, (Lee, 21)., CAT, CAT (H et l., 27). CAT H 2. 간조직내사멸화정도측정. epiderml growth ftor reeptor (EGFR) (Rper et l., 1987), mitogen-tivted protein kinse (MAPK) pthwy (Heo, 25; Jng nd Surh, 22). EGFR extrellulr signl-regulted kinses (ERK), p38, -Jun N-terminl kinses (JNK) Western lot., EGFR phospho-erk, SJE., phospho-jnk phosphop38, SJE (Fig. 5).,, MAPK pthwy (Sno et l., 21). MAPK pthwy 3 kinse, ERK, JNK p38 kinse, EGFR CAT tivity (%) p-erk p-jnk p-p38 GAPDH Experimentl group Fig. 4. Effets of SJE on hepti tlse (CAT) tivities in - indued rt. is ontrol group without dministrtion. (distilled wter 1.5 ml/rt) nd, 3 (S. jponi Extrt 15, 3 mg/kg rt, i.g) group were orl dministrted to rt for 1 dys efore tretment. Different supersripts represent signifintly different results (P<.5). Experimentl group Fig. 5. Chnges of liver toxiity relted protein expressions in -indued rts. The protein levels of EGFR, p-erk, p-jnk nd p-p38 were exmined fter tretment with or SJE (S. jponi Extrt).

6 다시마추출물의간보호효과 29. ERK, JNK p38 (Gujrl et l., 21; Kolh, 25). EGFR phospho-erk, JNK p38. SJE EGFR phospho- ERK, p38 JNK., SJE EGFR MAPK., GOT GPT., GSH, SOD, CAT., EGFR phospho-erk, phospho-jnk phospho-p38.,. 사사 (213 ). Referenes Blze T, Murry TK, MLughln JM nd Grie HC Hepti tests in toxiity studies on rts. Toxiol Appl Phrmol 3, Crpo HC, MCord MJ nd Fridovih I Preprtion nd ssy of superoxide dismutse. In Methods in enzymology, Fleisher, S. nd Pker, I. (eds.), Ademi Press, New York, U.S.A., 52, Fridovih I The iology of oxygen rdil. Siene 21, Gru S, Persohn E, Trommer WE nd Wolf A. 2. Mehnisms of ylosprine A-indued poptosis in rt heptoyte primry ultures. Toxiol Appl phrmol 163, Gujrl JS, Bui TJ, Frhood A nd Jeshke H. 21. Mehnism of ell deth during hepti ishemi-reperfusion in rts: Apoptosis or nerosis? Heptology 33, H KT, Kim YM, Kim CH, Kim DW, Choi DY nd Kim JK. 27. Study on the protetive effet of orni frutus ginst free rdil medited liver dmge. Koren J Orient Physiol Pthol 21, Hn SH, Kim JB, Min SG nd Lee CH The effets of Puerrie Rdix tehins dministrtion on liver funtion in ron tetrhloride-treted rts. J Koren So Food Nutr 24, Hng D, Choi HS, Kng SC, Kim KR, Sohn ES nd Kim MH. 25. Effets of fuoidn on NO prodution nd phgoytosis of mrophges nd the prolifertion of neuron ells. J Food Si Nutr 1, Hroun-Bouhedj F, Ellouli M, Sinquin C nd Boisson-Vidl C. 2. Reltionship etween sulfte groups nd iologil tivities of funs. Thromosis Res 1, Hyes JD nd Strnge RC. 2. Glutthione S-trnsferse polymorphisms nd their iologil onsequenes. Phrmology 61, Heo YJ. 25. Moleulr role of HBx nd AFT2 on ROSindued poptosis in humn liver ells. Mster s Thesis, Busn Ntionl University. Busn, Kore. Hidk I, Hino K, Koreng M, Gondo T, Nishin S, Ando M, Okud M nd Skid I. 27. Stronger Neo-minophgen C, glyyrrhizin-ontining preprtion, protets liver ginst ron tetrhloride-indued oxidtive stress in trnsgeni mie expressing the heptitis C virus polyprotein. Liver Int 27, Jng JH nd Surh YJ. 22. Bet-myloid indues oxidtive DNA dmge nd ell deth through tivtion of -Jun N terminl kinse. Ann N Y Ad Si 973, dx.doi.org/1.1111/j t4639.x. Jung SH, Lee YS, Lim SS, Lee S, Shin KH nd Kim, YS. 24. Antioxidnt tivities of isoflvones from the Rhizomes of Belmnd hinensis on ron tetrhloride-indued hepti injury in rts. Arh Phrm Res 27, Kim HK, Li L, Lee HS, Prk MO, Bilehl D, Li W nd Kim YH. 29. Protetive effets of Chlorell vulgris Extrt on ron tetrhloride-indued ute liver injury in mie. Food Si Biotehnol 18, Kolh W. 25. Coordinting ERK/MAPK signlling through sffolds nd inhiitors. Nt Rev Mol Cell Biol 6, Lee DS, Kim HR, Cho DM, Nm TJ nd Pyeun JH Uronte ompositions of lgintes from the edile rown lge. J Koren Fish So 31, 1-7. Lee MY. 21. Inhiitory Effet of Leek Green Juie on - indued Heptotoxiity in Rts. J Koren So Food Si Nutr 3, Lee TJ. 21. A study on liver funtion nd histologil hnges fter medition of llium stivum wter extrt in liver-dmged rts, PhD Thesis, Keimyung University, Degu, Kore. Nm CS, Kng KS nd H BJ. 27. Ulv ltu fuoidn extrt nd its protetive effets on -indued liver dysfuntion. Koren J Biotehnol Bioeng 22, Nishino T, Alzu Y nd Ngumo T The reltion etween

7 21 김미경ㆍ김인혜ㆍ남택정 the moleulr weight nd the ntiogulnt tivity of two types of fun sulftes from rown seweed Ekloni kurome. Agri Bio Chem 55, Prk HJ, Choi SC, Kim SH, Shin YH, Prk HJ nd Choi J. 25. Preventive effet of Shizndrin C derivtive DDB-mixed preprtion (DWP-4) ginst heptotoxiity indued y ron tetrhloride. Kor J Phrmogn 36, Prk PS, Lee BL nd Lee MY Effets of onion diet on ron tetrhloride toxiity of rts. J Koren So Food Nutr 2, Rper SE, Burwen SJ, Brker ME nd Jones AL Trnslotion of epiderml growth ftor to the heptoyte nuleus during rt liver regenertion. Gstroenterology 92, Rekngel RO Cron tetrhloride heptotoxiity. Phrmol Rev 19, Reitmn S nd Flnkel S A olorimetri method for the determintion of serum glutmi oxloeti nd glutmi pyruvi trnsminse. Am J Clin pthol 28, Roins B The sorption, distriution nd exretion of ron tetrhloride in dogs under vrious onditions. J Phrmol Exp Ther 37, Sno M, Fukud K, Sto T, Kwguhi H, Suemtsu M, Mtsud S, Koysu S, Mtsui H, Ymuhi-Tkihr K, Hrd M, Sito Y nd Ogw S. 21. ERK nd p38 MAPK, ut not NF-kpp B, re ritilly involved in retive oxygen speies-medited indution of IL-6 y ngiotensinⅡ in rdi firolsts. Cir Res 89, So MJ, Kim BK, Choi MJ, Kim KY, Rhee SH nd Cho EJ. 27. Protetive tivity of fuoidn nd lgini id ginst free rdil-indued oxidtive stress under in vitro nd ellulr system. J Food Si Nutr 12, Sunde RA nd Hoekstr WG Struture, synthesis nd funtion of glutthione peroxidse. Nutr Rev 38, Vendemile G, Altomre E, Grttglino I nd Alno O Inresed plsm levels of glutthione mlondildehyde fter ute ethnol ingestion in humns. J Heptol 9,

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DBPIA-NURIMEDIA J Koren So Food Si Nutr 한국식품영양과학회지 39(3), 33~39(2010) DOI: 10.376/jkfn.2010.39.3.33 녹차추출물이에탄올투여에의한초기간손상에미치는영향 김동춘 2 정승욱 2 박평심 1 1 조선대학교의학연구원 2 조선대학교의과대학생화학교실 Effets of Green Te Extrt on Aute -indued Heptotoxiity