多氯联苯暴露对食蚊鱼CYP19ɑ 和VTGɑ 基因的表达及骨骼形态雄性化的影响Effects of PCBs Exposure on CYP19ɑ ,VTGɑ Gene Expression and Morphological Masculinizing of Bone Morphogenesis in Mosquitofish( Gambusia affinis)
闫月明;方展强;
摘要(Abstract):
应用食蚊鱼细胞色素P450芳香化酶基因(CYP19%)和卵黄蛋白原基因(VTG%)m RNA转录水平为指标,研究微量多氯联苯(PCBs)长时间暴露对成年雌性食蚊鱼CYP19%基因和VTG%基因表达的影响,并评价其对雌性食蚊鱼产生的形态雄性化效应。采用静水暴露实验模式,分别设置0.08、0.4、2和10μg·L-1PCBs浓度,并设置对照组和平行组,定量测定14 d和28 d性腺CYP19%和肝脏VTG%m RNA表达水平的变化,以及对椎体脉棘发育的影响。暴露实验结果显示:1各PCBs浓度组(0.08、0.4、2和10μg·L-1PCBs)暴露均能抑制食蚊鱼CYP19%和VTG基因的表达,表明PCBs对食蚊鱼VTG%基因的抑制远大于对食蚊鱼CYP19%基因的抑制;20.4μg·L-1PCBs暴露显著地降低了食蚊鱼第15椎体脉棘的长度并降低第15、16椎体脉棘的L/D值,显示食蚊鱼出现骨骼形态雄性化,表明一定浓度的PCBs暴露会表现出抗雌激素效应。
关键词(KeyWords): 多氯联苯(PCBs);食蚊鱼;CYP19α;VTGα;m RNA表达;椎体脉棘;形态雄性化
基金项目(Foundation): 广东高校城市水环境生态治理与修复工程技术研究中心建设项目(No.2012gezx A004)
作者(Author): 闫月明;方展强;
Email:
DOI:
参考文献(References):
- [1]冯精兰,刘相甫,李怡帆,等.多氯联苯在我国环境介质中的分布[J].人民黄河,2011,33(2):86-89Feng J L,Liu X P,Li Y F,et al.The distribution of PCBs in China's environmental medium[J].Yel ow River,2011,33(2):86-89(in Chinese)
- [2]杜克久,储少岗,徐晓白,等.用MCF-7细胞增殖法研究国产多氯联苯的类雌激素效应[J].环境科学学报,2000,20(2):230-233Du K J,Chu S G,Xu X B,et al.Studies on the estrogenic effect of Chinese commercial PCBs by MCF-7 cell proliferation bioassay[J].Acta Scientiae Circumstantiae,2000,20(2):230-233(in Chinese)
- [3]杨丽丽,张晶,方展强.雌二醇、壬基酚、多氯联苯、镉和锌及其混合物对唐鱼的雌激素效应比较研究[J].水产学报,2011,35(6):102-109Yang L L,Zhang J,Fang Z Q.Comparative study of estrogenic effects of estradiol,nonylphenol,polychlorinated biphenyls,cadmium,zinc and its mixtures on Tanichthys albonubes[J].Journal of Fisheries of China,2011,35(6):102-109(in Chinese)
- [4]Vega-López A,Martínez-Tabche L,Domínguez-López ML,et al.Vitel ogenin induction in the endangered goodeid fish Girardinichthys viviparus:Vitel ogenin characterization and estrogenic effects of polychlorinated biphenyls[J].Comparative Biochemistry and Physiology,Part C,2006,142(3-4):356-364
- [5]CalòM,Alberghina D,Bitto A,et al.Estrogenic followed by anti-estrogenic effects of PCBs exposure in juvenil fish(Spaurus aurata)[J].Food and Chemical Toxicology,2010,48(8-9):2458-2463
- [6]Greytak S R,Champlin D,Cal ard G V.Isolation and characterization of two cytochrome P450 aromataseforms in kil ifish(Fundulus heteroclitus):Differential expressionin fish from pol uted and unpol uted environments[J].Aquatic Toxicology,2005,71(4):371–389
- [7]Kazeto Y,Place A R,Trant J M.Effects of endocrine disrupting chemicals on the expression of CYP19 genes in zebrafish(Danio rerio)juveniles[J].Aquatic Toxicology,2004,69(1):25-34
- [8]Orrego R,Mc Master M,Van Der Kraak G,et al.Effects of pulp and paper mil effluent extractives on aromatase CYP19a gene expression and sex steroid levels in juvenile triploid rainbow trout[J].Aquatic Toxicology,2010,97(4):353-360
- [9]Halm S,Pounds N,Maddix S.Exposure to exogenous 17beta-oestradiol disrupts P450arom B m RNA expression in the brain and gonad of adult fathead minnows(Pimephales promelas)[J].Aquatic Toxicology,2002,60(3-4):285-299
- [10]Contractor R G,Foran C M,Li S F,et al.Evidence of gender-and tissue-specific promoter methylation and the potential for ethinylestradiol-induced changes in Japanese medaka(Oryzias latipes)estrogen receptor and aromatase genes[J].Journal of Toxicology and Environmental Health Part A,2004,67(1):1-22
- [11]Kishida M,Mc Lel an M,Miranda J A,et al.Estrogen and xenoestrogens upregulate the brain aromatase isoform(P450arom B)and perturb markers of early development in zebrafish(Danio rerio)[J].Comparative Biochemistry and Physiology B-Biochemistry&Molecular Biology,2000,129(2-3):261-268
- [12]Rosen D E,Gordon M.Functional anatomy and evolution of male genitalia in Poeciliid fishes[J].Zoologica,1953,38(1):1-52
- [13]范俊杰,黎紫兰,徐少群,等.孕激素暴露致食蚊鱼形态雄性化的生物学效应[J].中国实验动物学报,2014,22(4):31-36Fan J J,Li Z L,Xu S Q,et al.The biological effects of morphological masculinization in mosquitofish(Gambusia affinis)exposed to progesterone[J].Acta Laboratorium Animalis Scientia Sinica,2014,22(4):31-36(in Chinese)
- [14]谢勇平,方展强.利用食蚊鱼目标基因转录水平评价东莞寒溪河雌/雄激素物质污染现状[J].水生生物学报,2013,37(4):691-697Xie Y P,Fang Z Q.Evaluation of estrogen/androgen pol ution by using the mosquitofish target gene expression in Hanxi River,Dongguan City[J].Acta Hydrobiologica Sinica,2013,37(4):1-7(in Chinese)
- [15]谢勇平,方展强.城市废水暴露致食蚊鱼形态雌/雄性化的生物学效应[J].生物灾害科学,2015,38(2):106-113Xie Y P,Fang Z Q.Urban sewage induces morphological feminization/masculinization in mosquitofish(Gambusia affinis)[J].Biological Disaster Science,2015,38(2):106-113(in Chinese)
- [16]侯丽萍,舒琥,李强,等.利用食蚊鱼目标基因转录水平评价四会市邓村河雌/雄激素物质污染现状[J].安徽农业大学学报,2015,42(1):152-158Hou L P,Shu H,Li Q,et al.Evaluation of estrogen/androgen pol ution by using the mosqueitofish target gene expression in Dengcun River,Sihui City[J].Journal of Anhui Agricultural University,2015,42(1):152-158(in Chinese)
- [17]Leusch F D L,Vanden Heuvel M R,Laurie A D,et al.Quantification of vitel ogenin m RNA induction in mosquitofish(Gambusia affinis)by reverse transcription real-time polymerase chain reaction(RT-PCR)[J].Biomarkers,2005,10(6):429-438
- [18]贺争鸣,李冠民,邢瑞昌.3R理论的形成、发展及在生命科学研究中的应用[J].实验动物科学与管理,2000,17(3):43-47He Z M,Li G M,Xing R C.The formation,development and application of 3R theory[J].Laboratory Animal Science&Administration,2000,17(3):43-47(in Chinese)
- [19]Rawson C,Lim R P,Warne M S J,et al.The effect of17β-estradiol on the development of modified hemal spines in early life-stage Gambusia holbrooki[J].Archives of Environmental Contamination and Toxicology,2006,51(2):253-262
- [20]Li L A.Polychlorinated biphenyl exposure and CYP19gene regulation in testicular and adrenocortical cell lines[J].Toxicology in Vitro,2007,21(6):1087-1094
- [21]Gregoraszczuk E L,Sowa M,Kajta M,et al.Effect of PCB 126 and PCB 153 on incidence of apoptosis in cultured theca and granulosa cells collected from small,medium and large preovulatory follicles[J].Reproductive Toxicology,2003,17(4):465-471
- [22]Wojtowicz A K,Goch M,Gregoraszczuk E L.Polychlorinated biphenyls(PCB 153 and PCB 126)action on conversion of 20-hydroxylated cholesterol to progesterone,androstenedione to testosterone,and testosterone to estradiol17beta[J].Experimental and Clinical Endocrinology&Diabetes,2005,113(8):464-470
- [23]Safe S H.Polychlorinated biphenyls(PCBs):Environmental impact,biochemical and toxic responses,and implications for risk assessment[J].Critical Review in Toxicology,1994,24(2):87-149
- [24]Mimura J,Fujii-Kuriyama Y.Functional role of Ah R in the expression of toxic effects by TCDD[J].Biochimica et Biophysica Acta,2003,1619:263-268
- [25]Bonefeld-Jorgensen E C,Andersen H R,Rasmussen T H,et al.Effect of highly bioaccumulated polychlorinated biphenyl congeners on estrogen and androgen receptor activity[J].Toxicology,2001,158(3):141-153
- [26]张伟,刘丛强,梁小兵,等.多氯联苯对小鼠肝微粒体CYP1A1酶活力及其m RNA表达的影响[J].毒理学杂志,2007,21(3):255-260Zhang W,Liu C Q,Liang X B,et al.Effects of PCBs on the activity of CYP1A1 and m RNA expression in mouse liver microsomal[J].Journal of Toxicology,2007,21(3):255-260(in Chinese)
- [27]Hou L,Xie Y,Ying G,et al.Developmental and reproductive characteristics of western mosquitofish(Gambusia affinis)exposed to paper mil effluent in the Dengcun River,Sihui,South China[J].Aquatic Toxicology,2011,103(3-4):140-149
- [28]Xie Y P,Fang Z Q,Hou L P,et al.Altered development and reproduction in western mosquitofish(Gambusia affinis)found in the Hanxi River,Southern China[J].Environmetal Toxicology and Chemistry,2010,29(11):2607-2615
- [29]闫月明.多氯联苯(Aroclor 1248)暴露对食蚊鱼CYP19αm RNA表达的影响及毒性效应研究[D].广州:华南师范大学,2013Yan Y M.Effects of PCBs(Aroclor 1248)exposure on the expression of gonadal cytochmme P450 aromatase(CYP19a)gene and the toxic effect in mosquitofish(Gambusia affinis)[D].Guangzhou:South China Normal University,2013(in Chinese)
- [30]Orlando E F,Bass D E,Caltabiano L M,et al.Altered development and reproduction in mosquitofish exposed to pulp and paper mil effluent in the Fenholloway River,Florida USA[J].Aquatic Toxicology,2007,84(3):399-405
- [31]Angus R A,Weaver S A,Grizzle J M,et al.Reproductive characteristics of male mosquitofish(Gambusia affinis)inhabiting a small southeastern US river receiving treated domestic sewage effluent[J].Environmental Toxicology and Chemistry,2002,21(7):1404-1409
- [32]范俊杰.雄激素和孕激素暴露对雌性食蚊鱼形态雄性化和生殖基因的影响[D].广州:华南师范大学,2012Fan J J.Vitel ogenin gene expression and morphological masculinization in female western mosquitofish(Gambusia affinis)exposed to androgens and progesterone[D].Guangzhou:South China Normal University,2012(in Chinese)
- [33]Rawson C A,Lim R P,Warne M St J,et al.The effect of17β-estradiol on the development of modified hemal spines in early life-stage Gambusia holbroohi[J].Archives of Environmental Contamination and Toxicology,2006,51(2):253-262
- [34]Rawson C A,Lim R P,Warne M St J.Skeletal morphology and maturation of male Gambusia holbroohi exposed to sewage treatment plant effluent[J].Ecotoxicology and Environmental Safety,2008,70(3):453-461
- [35]张晓婵,温茹淑,方展强.广州河涌污水暴露致食蚊鱼骨骼形态雄性化的影响[J].动物学杂志,2015,50(6):895-912Zhang X C,Wen R S,Fang Z Q.Morphological masculinizing effects of bone morphogenetic in mosquitofish(Gambusia affinis)exposed to municipal wastewater in Guangzhou creek[J].Chinese Journal of Zoology,2015,50(6):895-912(in Chinese)