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 68-737, 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 - ) http://dx.doi.org/1.5657/kfas.214.24 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 (http://retiveommons.org/lienses/y-n/3./)whih 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: +82. 51. 629. 5846 Fx: +82. 51. 629. 5842 E-mil ddress: nmtj@pknu..kr Copyright 214 The Koren Soiety of Fisheries nd Aquti Siene 24 pissn:374-8111, eissn:2287-8815
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, 3 3 95% 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 7. 4 1. () ( ) mouse, 1 1.5 ml/rt. 15, 3 mg/kg B.W. 1. 3, 1 ml/kg B.W. 1 24. 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 37 5 1 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 4. 4 5 1, 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 4. 4 5 1, 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
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) 4. 4 5 1, 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) 2 15 1 5 6 4 2 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
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 : 6.8 2 1 GSH level (%) 15 1 5 SOD tivity (U/mL) 8 6 4 2 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).
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 (%) 15 12 9 6 3 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).
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