苯并[a]芘对黑鲷肝脏GST活性的影响及其与肝脏代谢酶和胆汁代谢产物之间的变化关系Effects of Benzoapyrene Exposure on Hepatic GST Activity in Black Porgy(Sparus macrocephalus)and Variation Relationships with Hepatic Metabolic Enzymes and Biliary Metabolites
穆景利;王新红;林建清;林小龙;洪华生;
摘要(Abstract)：
为探讨苯并[a]芘(B[a]P)暴露对鱼类的影响并筛选特异、敏感的生物标志物,研究了B[a]P对黑鲷(Sparus macrocephalus)肝脏谷胱甘肽-S-转移酶(GST)活性的影响,并分析了肝脏7-乙氧基异吩唑酮-脱乙基酶(EROD)、GST活性与胆汁中B[a]P典型代谢产物3-羟基-苯并[a]芘(3-OHB[a]P)3者之间及与B[a]P之间的剂量、时间-效应关系.结果显示:在B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露期间,肝脏GST活性随暴露时间总体呈"钟"形的变化趋势,在第2d时达到峰值;在第12h、1d和2d时,GST活性与B[a]P暴露浓度呈显著正相关(R12h=0.966(p<0.05)、R1d=0.953(p<0.05)、R2d=0.824(p<0.05)).与前期研究结果对比分析发现,B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露7d时,胆汁中3-OHB[a]P浓度与B[a]P暴露浓度呈显著的剂量-效应关系(R=0.943,p<0.05);黑鲷肝脏GST、EROD活性与3-OHB[a]P的对数浓度均呈显著正相关(RGST=0.740(p<0.05)、REROD=0.839(p<0.05));2.0μg·L-1B[a]P暴露后,黑鲷肝脏EROD、GST活性与3-OHB[a]P随暴露时间的变化趋势基本相同,但并不完全一致.鱼类肝脏EROD、GST与胆汁中B[a]P代谢产物3者之间的变化关系较为复杂,暴露浓度和时间是影响其变化的重要因素,肝脏GST和EROD活性比胆汁中代谢产物更易受其影响,在B[a]P的环境监测中代谢产物可能是一种更为可行的生物标志物.
关键词(KeyWords)： 代谢产物;谷胱甘肽-S-转移酶;7-乙氧基异吩唑酮-脱乙基酶;代谢机制;胆汁;3-羟基-苯并[a]芘;黑鲷
基金项目(Foundation): 国家“863”项目(No.2007AA09Z126);; 国家自然科学基金项目(No.40106012;40476048;20777060);; 海洋公益性行业科研专项经费(No.200805090);; 国家海洋局青年基金(No.2008613);; 国家海洋局近岸海域生态环境重点实验室基金(No.200712)
作者(Author): 穆景利;王新红;林建清;林小龙;洪华生;
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DOI:
参考文献(References)：
- Bradford M M.1976.A rapid and sensitive method for thequantitation of microgram quantities of protein utilizing theprinciple of protein-dye binding[J].Analytical Biochemistry,72:248-254
- Buhler D R,Wang-Buhler J L.1998.Rainbow trout cytochromeP450s:purification,molecular aspects,metabolic activity,inductionand role in environmental monitoring[J].Comparative Biochemistryand Physiology Part C:Pharmacology,Toxicology andEndocrinology,121(1-3):107-137
- Conney A H.1982.Induction of microsomal enzymes by foreignchemicals and carcinogenesis by polycyclic aromatic hydrocarbons:G.H.A.Clowes Memorial Lecture[J].Cancer Research,42(12):4875-4917
- Flowers L,Bleczinski W,Burczynski M E,Harvey R G,PenningT M.1996.Dispositon and biological activity of benzo(a)pyrene-7,8-dione.A genotoxic metabolite generated by dihydrodioldehydrogenase[J].Biochemistry,35(42):13664-13672
- Fuentes-Rios D,Orrego R,Rudolph A,Mendoza G,GaviláJ F,Barra R.2005.EROD activity and biliary fluorescence inSchroederichthys chilensis(Guichenot1848):Biomarkers of PAHexposure in coastal environments of the South Pacific Ocean[J].Chemosphere,61(2):192-199
- Gagnon M M,Holdway D A.2002.EROD activity,serum SDHand PAH biliary metabolites in sand flathead(Platycephalusbassensis)collected in Port Phillip Bay,Australia[J].MarinePollution Bulletin,44(3):230-237
- Gao D,Luo Y,Guevara D,Wang Y,Rui M,Goldwyn B,LuY,Smith E C A,Lebwohl M,Wei H.2005.Benzo[a]pyreneand its metabolites combined with ultraviolet A synergisticallyinduce8-hydroxy-2′-deoxyguanosine via reactive oxygen species[J].Free Radical Biology and Medicine,39(9):1177-1183
- Goksr A,Flin L.1992.The cytochrome P-450system in fish,aquatic toxicology and environmental monitoring[J].AquaticToxicology,22(4):287-311
- Gorbi S,Regoli F.2004.Induction of cytochrome P4501A andbiliary PAH metabolites in European eel Anguilla anguillaSeasonal,dose-and time-response[J].Marine EnvironmentalReseach,58(2-5):511-515
- Gravato C,Santos M A.2002.Liver phaseⅠand phaseⅡenzymatic induction and genotoxic responses of[beta]-naphthoflavone water-exposed sea bass[J].Ecotoxicology andEnvironmental Safety,52(1):62-68
- Gravato C,Santos M A.2003.Genotoxicity biomarkers’associationwith B(a)P biotransformation in Dicentrarchus labrax L.[J].Ecotoxicology and Environmental Safety,55(3):352-358
- Habig W H,Pabst M J,Jakoby W B.1974.Glutathione S-Transferases.The first enzymetic step in mercapturic acidformation[J].Biological chemical,249(22):7130-7139
- Hasspieler B M,Behar J V,Digiulio R T.1994.Glutathione-dependent defense in channel catfish(Ictalurus punctatus)andbrown bullhead(Ameriurus nebulosus)[J].Ecotoxicology andEnvironmental Safety,28(1):82-90
- Hosnedl T,Kocourek V,TomaniováM,Volka K.2003.1-Hydroxypyrene as a biomarker for fish exposure to polycyclicaromatic hydrocarbons[J].Bulletin of Environmental Containedand Toxicological,71(3):465-472
- Kirby M F,Matthiessen P,Neall P,Tylor T,Allchin C R,KellyC A,Maxwell D L,Thain J E.1999.Hepatic EROD activity inflounder(Platichthys flesus)as an indicator of contaminantexposure in English estuaries[J].Marine Pollution Bulletin,38(8):676-686
- Klumpp D W,Hong H S,Humphrey C,Wang X H,Codi S.2002a.Toxic contaminants and their biological effects in coastalwaters of Xiamen,China.I.Organic pollutants in mussel andfish tissues[J].Marine Pollution Bulletin,44(8):752-760
- Klumpp D W,Humphrey C,Hong H S,Feng T.2002b.Toxiccontaminants and their biological effects in coastal waters ofXiamen,China.Ⅱ.Biomarkers and embryo malformation ratesas indicators of pollution stress in fish[J].Marine PollutionBulletin,44(8):761-769
- Little P J,James M O,Pritchard J B,Bend J R.1984.Benzo(a)pyrene metabolism in hepatic microsomes from feral and3-methycholanthrene-treated southern flounder,Paralichthys lethostigma[J].Journal of Environmental Pathology,Toxicology andOncology,5(4-5):309-320
- Melikian A A,Bagheri K,Hecht S S,Hoffmann D.1989.Metabolism of benzo[a]pyrene and7[beta],8[alpha]-dihydroxy-9[alpha],10[alpha]-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene in lung and liver of newborn mice[J].Chemico-Biological Interactions,69(2-3):245-257
- Mu J L,Wang X H,Lin J Q.2007.Dose-and time-dependentformation of3-hydroxy-benzo(a)pyrene in the bile of blackporgy(Sparus macrocephalus)exposed to benzo(a)pyrene[J].Environmental Chemical,26(4):487-490(in Chinese)
- Mu J L,Wang X H,Wang S H,Lin J Q,Wang K J,Hong HS.2009.Study on the kinetics of hepatic EROD activities inblack porgy(Sparusmacro cephalus)exposed to benzo(a)pyrene[J].Marine Science,33(1):68-71(in Chinese)
- Nacci D E,Kohan M,Pelletier M,George E.2002.Effects ofbenzo[a]pyrene exposure on a fish population resistant to thetoxic effects of dioxin-like compounds[J].Aquatic Toxicology,57(4):203-215
- Ruddock P J,Bird D J,McCalley D V.2002.Bile metabolitesof polycyclic aromatic hydrocarbons in three species of fish fromthe Severn Estuary[J].Ecotoxicological Environmental Safety,51(2):97-105
- Stegeman J J,Lech J J.1991.Cytochrome P-450monooxygenasesystems in aquatic species:carcinogen metabolism and biomarkersfor carcinogen and pollutant exposure[J].Environmental HealthPerspectives,90:101-109
- Van der Oost R,Beyer J,Vermuelen N P E.2003.Fishbioaccumulation and biomarkers in environmental risk assessment:a review[J].Environmental Toxicology and Pharmacology,13:57-149
- Van Schanke A,Holtz F,Van der Meer J P,Boon J P,ArieseF,Stroomberg G,Van den Berg M,Everaarts J M.2001.Dose-and time-dependent formation of biliary benzo[a]pyrene metabolitesin the marine flatfish dab(Limanda limanda)[J].EnvironmentalToxicology and Chemistry,20(8):1641-1647
- Wang C G,Zheng Y X,Zheng W Y,Yu Q.2004.The effectof exposure of benzo(a)pyrene,pyrene and their mixture onhepatic glutathione-S-transferase activity in Mugil so-iuy[J].Marine Science,28(3):40-43(in Chinese)
- Wang Y,Zhang R H,Zuo Z H,Chen Y X,Wang C G.2008.Relation of hepatic EROD activity and cytochrome P4501A levelin Sebastiscus marmoratus exposed to benzo[a]pyrene[J].Journal of Environmental Sciences,20:101-104
- Whyte J J,Jung R E,Schmitt C J,Tillitt D E.2000.Ethoxyresorufin-O-deethylase(EROD)activity in fish as abiomarker of chemical exposure[J].Critical Reviews in Toxicology,30(4):347-570
- Willett K L,Gardinali P R,Lienesch L A,Di Giulio R T.2000.Comparative metabolism and excretion of benzo(a)pyrenein2species of ictalurid catfish[J].Toxicological Sciences,58(1):68-76
- Zheng R H,Zhao Y,Chen Y X,Zuo Z H,Wang C G.2007.Effects of tributyl tin on the metabolism of benzo(a)pyrene inSebastiscus marmoratus[J].Acta Oceanologica Sinica,29(4):165-169(in Chinese)
- Zhu S,Li L,Thornton C,Carvalho P,Avery B A,Willett KL.2008.Simultaneous determination of benzo[a]pyrene andeight of its metabolites in Fundulus heteroclitus bile using ultra-performance liquid chromatography with mass spectrometry[J].Journal of Chromatography B,863(1):141-149
- Zhu X S,Meng F P,Yang Z X.2003.Comparative study onsensitivities of brain acetylcholinesterase from five marine fishesto two organophosphorus pesticides[J].Shanghai EnvironmentalScience,27(3):567-571(in Chinese)
- 穆景利,王新红,林建清.2007.黑鲷(Sparus macrocephalus)暴露苯并[a]芘后胆汁中代谢产物3-羟基-苯并[a]芘的剂量与时间-效应关系研究[J].环境化学,26(4):487-490
- 穆景利,王新红,王淑红,林建清,王克坚,洪华生.2009.苯并[a]芘影响黑鲷肝脏EROD活性变化的动力学研究[J].海洋科学,33(1):68-71
- 王重刚,陈奕欣,郑微云,余群.2004.苯并(a)芘、芘及其混合物暴露对梭鱼肝脏谷胱甘肽硫转移酶活性的影响[J].海洋科学,28(3):40-43
- 郑榕辉,赵扬,陈奕欣,左正宏,王重刚.2007.三丁基锡对褐菖鲉(Sebastiscus marmoratus)体内苯并(a)芘代谢的影响[J].海洋学报,29(4):165-169
- 朱小山,孟范平,杨正先.2003.5种海鱼脑AChE对2种有机磷农药的敏感性比较[J].上海环境科学,27(3):567-571