蒋鹏;盛南;王建设;戴家银;

• 1:中国科学院动物研究所中国科学院动物生态与保护生物学重点实验室

摘要(Abstract)：

五氯酚(pentachlorophenol,PCP)是一种持久性有机污染物,广泛用于灭钉螺、木材防腐、除草剂等方面,由于PCP在环境中的持久性和生物累积性,其对生态环境和人类健康造成潜在危害。本文以鸡肝癌细胞系(chicken hepatoma cells,LMH)为受试对象,探讨了PCP对细胞色素P450(CYP450)和抗氧化系统的影响。MTT结果显示LMH细胞经不同浓度PCP暴露后,呈现出先促进细胞增殖后抑制的J-型曲线,PCP对LMH细胞24 h的半数效应浓度(24 h-EC50)为427.52μmol·L~(-1)。LMH细胞在1.56、6.25、25、100μmol·L~(-1)PCP染毒条件下可增加细胞EROD、MROD、PROD和BFC活性,并可使CYP1A、1B、1C、2H及3A家族基因mRNA表达水平升高。LMH细胞在0.4~100μmol·L~(-1)PCP染毒下可显著降低硫酸基转移酶(SULT1B1和SULT 1C1)基因mRNA水平。此外,LMH细胞在6.25、25、100μmol·L~(-1)PCP染毒下可引起细胞内ROS升高,同时PCP(1.56~100μmol·L~(-1))可显著增加细胞内MDA含量和降低GSH/GSSH比值。这些结果表明细胞色素P450(CYP450)基因及酶活性的变化、细胞内ROS和MDA含量及GSH/GSSH可作为评价LMH细胞PCP毒性效应的敏感性生物标志物。此研究在细胞水平上利用多个评价指标研究PCP对细胞的毒性效应,为PCP环境风险评价提供依据。

关键词(KeyWords)： 五氯酚;鸡肝癌细胞系;细胞色素P450;硫酸基转移酶;氧化应激

Abstract:

Keywords:

基金项目(Foundation): 广东省自然科学基金(2015A030312005)

作者(Author): 蒋鹏;盛南;王建设;戴家银;

Email:

参考文献(References)：

• [1]Crosby D G.IUPAC reports on pesticides.14.Environmental chemistry of pentachlorophenol[J].Pure and Applied Chemistry,1981,53:1051-1080
• [2]Okeke B C,Paterson A,Smith J E,et al.Comparative biotransformation of pentachlorophenol in soils by solid substrate cultures of Lentinula edodes[J].Applied Microbiology and Biotechnology,1997,48:563-569
• [3]Zheng W W,Yu H,Wang X,et al.Systematic review of pentachlorophenol occurrence in the environment and in humans in China:Not a negligible health risk due to the re-emergence of schistosomiasis[J].Environment International,2012,42:105-116
• [4]Jorens P G,Schepens P J C.Human pentachlorophenol poisoning[J].Human&Experimental Toxicology,1993,12:479-495
• [5]Law W M,Lau W N,Lo K L,et al.Removal of biocide pentachlorophenol in water system by the spent mushroom compost of Pleurotus pulmonarius[J].Chemosphere,2003,52:1531-1537
• [6]Reigner B G,Bois F Y,Tozer T N.Assessment of pentachlorophenol exposure in humans using the clearance concept[J].Human&Experimental Toxicology,1992,11:17-26
• [7]Blakley B R,Yole M J,Brousseau P,et al.Effect of pentachlorophenol on immune function[J].Toxicology,1998,125:141-148
• [8]Chhabra R S,Maronpot R M,Bucher J R,et al.Toxicology and carcinogenesis studies of pentachlorophenol in rats[J].Toxicological Sciences,1999,48:14-20
• [9]Roberts H J.Pentachlorophenol-associated aplastic anemia,red cell aplasia,leukemia and other blood disorders[J].The Journal of the Florida Medical Association,1990,77:86-90
• [10]Umemura T,Kai S,Hasegawa R,et al.Pentachlorophenol(PCP)produces liver oxidative stress and promotes but does not initiate hepatocarcinogenesis in B6C3F1 mice[J].Carcinogenesis,1999,20:1115-1120
• [11]Zhu B Z,Shan G Q.Potential mechanism for pentachlorophenol-induced carcinogenicity:A novel mechanism for metal-independent production of hydroxyl radicals[J].Chemical Research in Toxicology,2009,22:969-977
• [12]Shan G Q,Ye M Q,Zhu B Z,et al.Enhanced cytotoxicity of pentachlorophenol by perfluorooctane sulfonate or perfluorooctanoic acid in Hep G2 cells[J].Chemosphere,2013,93:2101-2107
• [13]Westerink W M A,Schoonen W G E J.Cytochrome P450 enzyme levels in Hep G2 cells and cryopreserved primary human hepatocytes and their induction in Hep G2 cells[J].Toxicology in Vitro,2007,21:1581-1591
• [14]Dong H,Xu D M,Hu L H,et al.Evaluation of N-acetyl-cysteine against tetrachlorobenzoquinone-induced genotoxicity and oxidative stress in Hep G2 cells[J].Food and Chemical Toxicology,2014,64:291-297
• [15]Nguyen T N T,Bertagnolli A D,Villalta P W,et al.Characterization of a deoxyguanosine adduct of tetrachlorobenzoquinone:Dichlorobenzoquinone-1,N-2-etheno-2'-deoxyguanosine[J].Chemical Research in Toxicology,2005,18:1770-1776
• [16]孙毓鑫.鸟类作为指示生物监测陆生环境中卤代有机污染物的研究[D].北京:中国科学院研究生院,2012
• [17]Hu J Y,Li J,Wang J S,et al.Synergistic effects of perfluoroalkyl acids mixtures with J-shaped concentration-responses on viability of a human liver cell line[J].Chemosphere,2014,96:81-88
• [18]Dorsey W C,Tchounwou P B,Ford B D.Neuregulin 1-beta cytoprotective role in AML 12 mouse hepatocytes exposed to pentachlorophenol[J].International Journalof Environmental Research and Public Health,2006,3:11-22
• [19]Ling B,Gao B,Yang J.Evaluating the effects of tetrachloro-1,4-benzoquinone,an active metabolite of pentachlorophenol,on the growth of human breast cancer cells[J].Journal of Toxicology,2016(5):1-8
• [20]Dorsey W C,Tchounwou P B.CYP1a1,HSP70,P53,and c-fos expression in human liver carcinoma cells(Hep G2)exposed to pentachlorophenol[J].Biomedical Sciences Instrumentation,2003,39:389-396
• [21]金帮明,王辅明,熊力,等.五氯酚对Hela细胞毒性及DNA损伤的研究[J].环境科学,2012,33(2):658-664Jin B M,Wang F M,Xiong L,et al.Effects of pentachlorophenol on DNA damage and cytotoxicity of Hela cells[J].Environmental Science,2012,33(2):658-664(in Chinese)
• [22]Ibrahim Z S.Chenodeoxycholic acid increases the induction of CYP1A1 in Hep G2 and H4IIE cells[J].Experimental and Therapeutic Medicine,2015,10(5):1976-1982
• [23]Pelkonen O,Turpeinen M,Hakkola J,et al.Inhibition and induction of human cytochrome P450 enzymes:Current status[J].Archives of Toxicology,2008,82:667-715
• [24]He X T,Nie X P,Wang Z H,et al.Assessment of typical pollutants in waterborne by combining active biomonitoring and integrated biomarkers response[J].Chemosphere,2011,84:1422-1431
• [25]Marshall N B,Kerkvliet N I.Dioxin and immune regulation:Emerging role of aryl hydrocarbon receptor in the generation of regulatory T cells[J].Annals of the New York Academy of Sciences,2010,1183:25-37
• [26]Sorg O.AhR signalling and dioxin toxicity[J].Toxicology Letters,2014,230:225-233
• [27]Jung D K,Klaus T,Fent K.Cytochrome P450 induction by nitrated polycyclic aromatic hydrocarbons,azaarenes,and binary mixtures in fish hepatoma cell line PLHC-1[J].Environmental Toxicology and Chemistry,2001,20:149-159
• [28]Thummel K E,Wilkinson G R.In vitro and in vivo drug interactions involving human CYP3A[J].Annual Review of Pharmacology and Toxicology,1998,38:389-430
• [29]Mortensen A S,Arukwe A.Modulation of xenobiotic biotransformation system and hormonal responses in Atlantic salmon(Salmo salar)after exposure to tributyltin(TBT)[J].Comparative Biochemistry and Physiology.Toxicology&Pharmacology:CBP,2007,145:431-441
• [30]Hirakawa S,Iwata H,Takeshita Y,et al.Molecular characterization of cytochrome P450 1A1,1A2,and1B1,and effects of polychlorinated dibenzo-p-dioxin,dibenzofuran,and biphenyl congeners on their hepatic expression in Baikal seal(Pusa sibirica)[J].Toxicological Sciences,2007,97:318-335
• [31]Kubota A,Watanabe M X,Kim E Y,et al.Accumulation of dioxins and induction of cytochrome P450 1A4/1A5 enzyme activities in common cormorants from Lake Biwa,Japan:Temporal trends and validation of national regulation on dioxins emission[J].Environmental Pollution,2012,168:131-137
• [32]Esler D,Ballachey B E,Trust K A,et al.Cytochrome P4501A biomarker indication of the timeline of chronic exposure of Barrow's goldeneyes to residual Exxon Valdez oil[J].Marine Pollution Bulletin,2011,62:609-614
• [33]Bigorgne E,Custer T W,Dummer P M,et al.Chromosomal damage and EROD induction in tree swallows(Tachycineta bicolor)along the Upper Mississippi River,Minnesota,USA[J].Ecotoxicology,2015,24:1028-1039
• [34]Iwata H,Nagahama N,Kim E Y,et al.Effects of in ovo exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin on hepatic AHR/ARNT-CYP1A signaling pathways in common cormorants(Phalacrocorax carbo)[J].Comparative Biochemistry and Physiology.Toxicology&Pharmacology:CBP,2010,152:224-231
• [35]Honma W,Kamiyama Y,Yoshinari K,et al.Enzymatic characterization and interspecies difference of phenol sulfotransferases,ST1A forms[J].Drug Metabolism and Disposition:the Biological Fate of Chemicals,2001,29:274-281
• [36]Meloche C A,Falany C N.Expression and characterization of the human 3 beta-hydroxysteroid sulfotransferases(SULT2B1a and SULT2B1b)[J].The Journal of Steroid Biochemistry and Molecular Biology,2001,77:261-269
• [37]Mulder G J,Scholtens E.Phenol sulphotransferase and uridine diphosphate glucuronyltransferase from rat liver in vivo and vitro.2,6-Dichloro-4-nitrophenol as selective inhibitor of sulphation[J].The Biochemical Journal,1977,165:553-559
• [38]Koster H,Scholtens E,Mulder G J.Inhibition of sulfation of phenols in vivo by 2,6-dichloro-4-nitrophenol:Selectivity of its action in relation to other conjugations in the rat in vivo[J].Medical Biology,1979,57:340-344
• [39]Wang L Q,James M O.Inhibition of sulfotransferases by xenobiotics[J].Current Drug Metabolism,2006,7:83-104
• [40]Nagata K,Ozawa S,Miyata M,et al.Isolation and expression of a c DNA-encoding a male-specific rat sulfotransferase that catalyzes activation of N-hydroxy-2-acetylaminofluorene[J].Journal of Biological Chemistry,1993,268:24720-24725
• [41]D'Andre H C,Paul W,Shen X,et al.Identification and characterization of genes that control fat deposition in chickens[J].Journal of Animal Science and Biotechnology,2013,4(1):1-16
• [42]Wilson L A,Reyns G E,Darras V M,et al.c DNA cloning,functional expression,and characterization of chicken sulfotransferases belonging to the SULT1B and SULT1C families[J].Archives of Biochemistry and Biophysics,2004,428:64-72
• [43]Landry W D,Cotter T G.ROS signalling,NADPH oxidases and cancer[J].Biochemical Society Transactions,2014,42:934-938
• [44]Chen E I.Mitochondrial dysfunction and cancer metastasis[J].Journal of Bioenergetics and Biomembranes,2012,44:619-622
• [45]Jiang P,Wang J,Zhang J,et al.Effects of pentachlorophenol on the detoxification system in white-rumped munia(Lonchura striata)[J].Journal of Environmental Sciences,2016,44:224-234