Asian Journal of Ecotoxicology GST. 3 -OH B a P. R GST =0.740 p <0.05 R EROD =0.839 p < μg L -1 B a P X171.5

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1 Asian Journal of Ecotoxicology Vol 4, 2009 No 4, a GST * a B a P B a P Sparus macrocephalus -S- GST 7 - EROD GST B a P a 3 -OH B a P 3 B a P - B a P μg L -1 GST 2d 12h 1d 2d GST B a P R 12h =0966 p<005 R 1d = 0953 p<005 R 2d =0824 p<005 B a P μg L -1 7d 3 -OH B a P B a P - R=0943 p<005 GST EROD 3 -OH B a P R GST =0740 p <005 R EROD =0839 p <005 20μg L -1 B a P EROD GST 3 -OH B a P EROD GST B a P 3 GST EROD B a P -S a X1715 A Effects of Benzo a pyrene Exposure on Hepatic GST Activity in Black Porgy Sparus macrocephalus and Variation Relationships with Hepatic Metabolic Enzymes and Biliary Metabolites MU Jing-li 1 2 WANG Xin-hong 1 * LIN Jian-qing 3 LIN Xiao-long 1 HONG Hua -sheng 1 1 State Key Laboratory of Marine Environmental Science, Environmental Science Research Center, Xiamen University, Xiamen National Marine Environmental Monitoring Center, Dalian College of Bioengineering, Jimei University, Xiamen Received 13 December 2008 accepted 4 January 2009 Abstract Effects of benzo a pyrene B a P exposure on the activity of glutathione S -transferase GST in black porgy Sparus macrocephalus liver was studied The dose and time response of two enzymes 7 -ethoxyresorufin O -deethylase EROD and GST involved in B a P metabolism and the amounts of B a P metabolites excreted into the bile were investigated Results showed that the time -response trend on of induction of hepatic GST of black porgy was bell -shaped with a plateau at B a P exposure for 2 days A significant dose -related increase of the GST activity in liver was observed at the exposure time of 12h, 1d and 2d R 12h =0966, p<005; R 1d =09529, p<005; R 2d =0824, p<005 Compared with our former study, the induction of hepatic EROD, GST activities and biliary 3 -OH B a P of black porgy were significantly induced by B a P 05, 10, 20 and 50μg L -1 after exposure for 7 days and a significant dose -response relationship between 3 -OH B a P and B a P was observed R=0943, p<005 After exposed to 20μg L -1 B a P, similar variation trends of hepatic EROD, GST activities and biliary 3-OH B a P had been observed in black porgy, but those variations were not consistency with the time change completely Those results demonstrated that the variation between hepatic metabolic enzymes and biliary metabolites were very complex The exposure concentrations and exposure time were the most important factors which influence the variations of metabolic enzymes and metabolites Keywords metabolites glutathione S -transferase 7 -ethoxyresorufin O -deethylase metabolic mechanism bile 3 -OH B a P Sparus macrocephalus No 2007AA09Z126 No No No No * Corresponding author xhwang@xmueducn

2 4 a GST Introduction a Benzo a pyrene B a P EROD Polycyclic aromatic PAHs 2007 hydrocarbons PAHs Sebastiscus marmoralus B a P Conney, 1982 Nacci et al, B a P B a P EROD B a P B a P B a P P4501A CYP1A B a P Sparus macrocephalus B a P Little et al, 1984 Melikian et al, 1989 Flowers et al, 1996 Gao et al, ethoxyresorufin O -deethylase EROD CYP1A1-2002b B a P EROD EROD B a P CYP4501A1 B a P a 3 -OH B a P 3 - Goks 覬 r and F 覬 lin, 1992 Buhler and Wang -Buhler, 1998 Whyte et al, S - Glutathione S - transferase GST Glutathione B a P GSH B a P Habig 2 Materials and methods et al, 1974 Hasspieler et al, 1994 Van Schanke et al, 2001 B a P 21 HPLC B a P Ruddock et al, 2002 Hosnedl et al, 2003 Zhu et al, 2008 EROD GST B a P >96% Sigma -Aldrich PAHs μg L -1 Gagnon and Holdway, L 2h 6h 12h 1d 2d 4d Gravato and Santos, 2002 Gagnon and Holdway, Klummp 2002b EROD GST 14d OH Py PAHs 7d Gorbi Regoli 2004 EROD Fuentes -Rios μg L -1 56d EROD GST EROD GST Klumpp et al, 2002a GST GST PAHs B a P 912±13 cm 1120±15 g 115cm 63cm 50cm 7d 14d 6

3 Results and analysis 30±15 mg 15mL ph=76 31 B a P GST 2min -4 B a P GST 10000r min -1 20min EROD 1 GST GST GST Habig GST 14d 01mL CDNB 175mL 2,4-37 1min GST 025mL 37 5min GSH GST 37 1min 1cm GSH B a P 340nm A1 37 3min A2 GST = A2 -A /Δt GST Bradford Stegeman and Lech, 1991 Hasspieler et al, 1994 Bradford, 1976 EROD B a P GST ± SPSS Pearson 2d B a P B a P 4d GST GSH B a P GST 2004 B a P 5μg L μg L -1 B a P GST ANOVA Tukey

4 4 a GST 519 5μg L -1 B a P 1d GST 32 EROD GST 3 -OH B a P 7d B a P EROD B B a P 3 -OH B a P a P - GST EROD GST 3-OH B a P 3 B a P 12h 1d 2d 2 B a P R 12h =0966 p <005 R 1d =0953 p <005 R 2d =0824 p<005 B a P 321 EROD GST 3 -OH B a P 3 GST B a P B a P - GST Hepatic GST activity/ nmol min -1 mg h 茛 1d 2d 茛茛茛茛 B a P B a P concentration/ μg L -1 茛 B a P CYP450 CYP1A B a P a -7,8 - -9,10 - BPDE B a P 3 -OH B a P a 9 -OH B a P GSH Little et al, 1984 Willett et al, 2000 EROD CYP1A B a P 2 GST B a P GST a R 12h=0966, p<005; R 1d=0952, p<005; R 2d=0824, p<005 a GSH Fig2 Variation of GST activity in liver of black porgy Habig et al, 1974 Kirby et al, 1999 exposed to B a P R 12h=0966, p<005; R 1d=0952, p<005; R 2d=0824, p< OH B a P B a P - GST 7d 3 -OH B a P Van der p<005 Oost EROD GST 33% 48% GST GST B a P GST 7d EROD GST 3 -OH B a P 0839 p<005 GST p < B a P

5 520 4 B a P EROD PAHs Van Schanke Gorbi Regoli Limanda limanda B a P Anguilla anguilla PAHs 7d B a P B a P EROD EROD GST B a P PAHs 3 B a P B a P EROD GST EROD B a P PAHs EROD GST EROD Wang 2008 PAHs B a P B a P B a P - 3 -OH B a P B a P B a P CYP1A B a P B a P PAHs 3-OH B a P 3-OH B a P concentration/ ng μl EROD Hepatic EROD activity/ pmol min -1 mg OH B a P EROD GST B a P B a P concentration/ μg L -1 3 B a P 7d EROD GST 3-OH B a P B a P - Fig3 Dose-effect relationships between B a P exposure concentrations and EROD, GST activities and 3-OH B a P concentration after exposure for 7 days GST Hepatic GST activity/ nmol min -1 mg -1 Log EROD EROD GST days units of 3-OH B a P, GST, EROD are ng μl -1, pmol min -1 mg -1, nmol min -1 mg-1, respectively Log 3-OH B a P 4 B a P 7d EROD GST 3-OH B a P 3-OH B a P GST EROD ng μl -1 pmol min -1 mg-1 nmol min -1 mg-1 Fig4 The correlativity between hepatic EROD, GST activities and 3-OH B a P concentration after B a P exposure for Log GST 322 EROD GST 3 -OH B a P 3-20μg L -1 B a P EROD GST 3-OH B a P 3 5 EROD GST 3 -OH B a P 2d GST EROD 3 -OH B a P 7d 3 - B a P CYP1A B a P B a P GST B a P 3 -OH B a P GSH

6 4 a GST 521 B a P B a P B a P B a P GSH GSH Habig et al, 1974 Conney, 1982 Whyte et al, 2000 B a P 2d B a P PAHs GST 3-OH B a P 5 EROD B a P GST - B a P B a P B a P Willett 2000 Ictalurus Punctatus B a P Dicentrarchus labrax 012μmol L -1 B a P B a P 3d GST EROD GST B a P B a P EROD GST GST B a P 4h 8h 16h Gravato Santos 05μg L -1 B a P 8 GST Gravato Santos h EROD 3-OH B a P 3-OH B a P concentration/ ng μl OH B a P a EROD GST 2h 6h 12h 1d 2d 4d 7d 14d 21d Time EROD Hepatic EROD activity/ pmol min -1 mg GST Hepatic GST activity/ nmol min -1 mg -1 5 Fig5 3-OH B a P 3-OH B a P concentration/ ng μl OH B a P EROD GST b 00 2h 6h 12h 1d 2d 4d 7d 14d 21d 0 20 Time 20μg L -1 B a P EROD GST 3-OH B a P a: 20μg L -1 b: Changes of hepatic EROD, GST activities and 3-OH B a P concentration with time under the exposure of 20μg L -1 B a P a: 20μg L -1 B a P; b: acetone control EROD Hepatic EROD activity/ pmol min -1 mg GST Hepatic GST activity/ nmol min -1 mg -1

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