慢性镉暴露对斑马鱼卵巢组织的氧化损伤和子代发育的母源影响
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摘要
目的:采用半静态毒性实验方法,研究慢性镉暴露对性成熟斑马鱼卵巢组织抗氧化酶活性、脂质过氧化水平的影响及其母源传递毒性效应。方法:依据前期测定的96 h LC50,参考国家渔业标准,设置5个镉处理组(0.058、0.116、0.232、0.580、1.160 mg/L)和1个对照组,镉暴露21 d后,对卵巢组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)活性及脂质过氧化物丙二醛(MDA)含量进行测定,并对其受精率、F1代胚胎孵化率及96 hpf幼鱼体长进行统计分析。结果:不同浓度镉暴露均导致卵巢组织SOD、CAT和GPx活性显著降低、MDA含量显著升高并呈浓度依赖效应。子代母源毒性分析显示,与对照组相比,0.232和0.580 mg/L浓度组受精率均显著下降,0.232 mg/L浓度组F1代胚胎孵化率显著降低;各浓度组受精后96 h幼鱼体长与对照组相比有显著性差异,且随着镉浓度的增加体长显著变短。结论:慢性镉暴露导致斑马鱼卵巢组织抗氧化酶活性受到显著抑制,对组织和细胞造成严重氧化损伤,并通过母源传递导致子代发育的毒性效应。
Objective: The effects of chronic cadium(Cd) exposure on the activities of antioxidant enzymes and the level of lipid peroxidation in ovaries of mature zebrafish, as well as the maternal toxicity effects on offspring were studied by using semi-static toxic test method. Methods: According to 96 h LC50 values and the National Fisheries Standards of China, we set five Cd treatment groups(0.058, 0.116, 0.232, 0.580, 1.160 mg/L) and one control group without Cd. After Cd exposure for 21 d, the activities of superoxide dismutase(SOD), catalase(CAT)and glutathione peroxidase(GPx), and the level of malonyldialdehyde(MDA) in zebrafish ovaries were investigated.Moreover, the fertilization success rate, hatching rate and 96 hpf-body length of their offspring(F1 larvae) were studied. Results: Compared with those in the control group, the activities of antioxidant enzymes SOD, CAT and GPx were decreased significantly, and the content of MDA was increased significantly in each Cd-treated group with a concentration-dependent manner. The fertilization success rates were reduced significantly in the 0.232 and0.580 mg/L Cd-treated groups and the larvae hatching rate was decreased significantly in the 0.232 mg/L Cd-treated group compared with that in the control group. The body length was shorter in each Cd-treated group compared with that in the control group. Conclusion: These data indicated that the activities of antioxidant enzymes were significantly inhibited after Cd exposure for 21 d, resulting in severe oxidative damage in ovary of mature zebrafish. The exposure history of mothers caused toxic effects on the fertilization and hatching as well as growth of the next non-exposed generation of embryos and larvae.
Objective: The effects of chronic cadium(Cd) exposure on the activities of antioxidant enzymes and the level of lipid peroxidation in ovaries of mature zebrafish, as well as the maternal toxicity effects on offspring were studied by using semi-static toxic test method. Methods: According to 96 h LC50 values and the National Fisheries Standards of China, we set five Cd treatment groups(0.058, 0.116, 0.232, 0.580, 1.160 mg/L) and one control group without Cd. After Cd exposure for 21 d, the activities of superoxide dismutase(SOD), catalase(CAT)and glutathione peroxidase(GPx), and the level of malonyldialdehyde(MDA) in zebrafish ovaries were investigated.Moreover, the fertilization success rate, hatching rate and 96 hpf-body length of their offspring(F1 larvae) were studied. Results: Compared with those in the control group, the activities of antioxidant enzymes SOD, CAT and GPx were decreased significantly, and the content of MDA was increased significantly in each Cd-treated group with a concentration-dependent manner. The fertilization success rates were reduced significantly in the 0.232 and0.580 mg/L Cd-treated groups and the larvae hatching rate was decreased significantly in the 0.232 mg/L Cd-treated group compared with that in the control group. The body length was shorter in each Cd-treated group compared with that in the control group. Conclusion: These data indicated that the activities of antioxidant enzymes were significantly inhibited after Cd exposure for 21 d, resulting in severe oxidative damage in ovary of mature zebrafish. The exposure history of mothers caused toxic effects on the fertilization and hatching as well as growth of the next non-exposed generation of embryos and larvae.
引文
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[12]马丹旦,雷雯雯,吴昊,等.急性镉染毒对河南华溪蟹精子质量的影响[J].环境科学学报,2013,33(7):2044-2049.
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[14]Simon O,Floc'H E,Geffroy B,et al.Exploring ecotoxicological fish bioassay for the evaluation of uranium reprotoxicity[J].Environ Toxicol Chem,2014,33(8):1817-1824.
[15]刘燕.水体中镉暴露对稀有鮈鲫氧化应激及组织学影响[D].重庆:西南大学,2014:17-25.
[16]Adeogun A O,Onibonoje K,Ibor O R,et al.Endocrine-disruptor molecular responses,occurrence of intersex and gonado-histopathological changes in tilapia species from a tropical freshwater dam(Awba Dam)in Ibadan,Nigeria[J].Aquat Toxicol,2016,174:10-21.
[17]Chouchene L,Banni M,Kerkeni A,et al.Cadmiuminduced ovarian pathophysiology is mediated by change in gene expression pattern of zinc transporters in zebrafish(Danio rerio)[J].Chem Biol Interact,2011,193(2):172-179.
[18]Wang Y X,Ferrari M C O,Hoover Z,et al.The effects of chronic exposure to environmentally relevant levels of waterborne cadmium on reproductive capacity and behaviour in fathead minnows[J].Arch Environ Contam Toxicol,2014,67(2):181-191.
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[20]Kusch R C,Krone P H,Chivers D P.Chronic exposure to low concentrations of waterborne cadmium during embryonic and larval development results in the long-term hindrance of antipredator behavior in zebrafish[J].Environ Toxicol Chem,2008,27(3):705-710.
[21]Vaziri N D,Lin C Y,Farmand F,et al.Superoxide dismutase,catalase,glutathione peroxidase and NADPH oxidase in lead-induced hypertension[J].Kidney Int,2003,63(1):186-194.
[22]Roméo M,Bennani N,Gnassia-Barelli M,et al.Cadmium and copper display different responses towards oxidative stress in the kidney of the sea bass Dicentrarchus labrax[J].Aquat Toxicol,2000,48(2-3):185-194.
[23]Banni M,Chouchene L,Said K,et al.Mechanisms underlying the protective effect of zinc and selenium against cadmium-induced oxidative stress in zebrafish Danio rerio[J].Biometals,2011,24(6):981-992.
[24]李文英,刘荣,蒋园,等.DBP对斑马鱼肝脏和鳃SOD及ATPase酶活性的影响[J].水生态学杂志,2007,27(4):15-18.
[25]熊道文,方涛,陈旭东,等.纳米材料对斑马鱼的氧化损伤及应激效应研究[J].环境科学,2010,31(5):1320-1327.
[26]闫博,王兰,李涌泉,等.镉对长江华溪蟹肝胰腺抗氧化酶活力的影响[J].动物学报,2007,53(6):1121-1128.
[27]马京津,徐团,轩瑞晶,等.铅对河南华溪蟹卵巢抗氧化酶活力和脂质过氧化水平的影响[J].环境科学学报,2012,32(9):2346-2352.
[28]徐团,李颖君,金芬芬.镉对长江华溪蟹精巢抗氧化酶活性及脂质过氧化的影响[J].四川动物,2011,30(5):762-764.
[29]刘娜,闫博,李涌泉,等.Cd2+对长江华溪蟹谷胱甘肽系统的影响[J].环境科学,2008,29(8):2302-2307.
[30]Iszard M B,Liu J,Klaassen C D.Effect of several metallothionein inducers on oxidative stress defense mechanisms in rats[J].Toxicology,1995,104(1-3):25-33.
[31]Zheng J L,Yuan S S,Wu C W,et al.Acute exposure to waterborne cadmium induced oxidative stress and immunotoxicity in the brain,ovary and liver of zebrafish(Danio rerio)[J].Aquat Toxicol,2016,180:36-44.
[32]Su M W,Pei R T,Yan C J.Maternal cadmium exposure induces mt2,and smt B,m RNA expression in zebrafish(Danio rerio)females and their offspring[J].Comp Biochem Physiol Cy,2012,156(156):1-6.
[33]Sellin M K,Kolok A S.Cadmium exposures during early development:Do they lead to reproductive impairment in fathead minnows[J]?Environ Toxicol Chem,2006b,25(11):2957-2963.
[34]Bourrachot S,Brion F,Pereira S,et al.Effects of depleted uranium on the reproductive success and F1generation survival of zebrafish(Danio rerio)[J].Aquat Toxicol,2014,154(1):1-11.
[35]Sellin M K,Eidem T M,Kolok A S.Cadmium exposures in fathead minnows:are there sex-specific differences in mortality,reproductive success,and Cd accumulation[J]?Arch Environ Contam Toxicol,2007,52(4):535-540.
[36]Sellin M K,Kolok A S.Cd exposures in fathead minnows:effects on adult spawning success and reproductive physiology[J].Arch Environ Contam Toxicol,2006a,51(4):594-599.
[37]Shi X,Du Y,Lam P K S,et al.Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS[J].Toxicol Appl Pharmacol,2008,230(1):23-32.
[38]鲁疆,王占洋,袁玉婷,等.氯化镉对斑马鱼胚胎的发育毒性[J].生态毒理学报,2013,8(3):381-388.
[39]Zhu B,Lei L,Li D L,et al.Developmental toxicity in rare minnow(Gobiocypris rarus)embryos exposed to Cu,Zn and Cd[J].Ecotoxicol Environ Safety,2014,104C(1):269-277.
[2]张翠,翟毓秀,宁劲松,等.镉在水生动物体内的研究概况[J].水产科学,2007,26(8):465-470.
[3]Elsayed Y S,Elgazzar A M,Elnahas A F,et al.Vitamin C modulates cadmium-induced hepatic antioxidants'gene transcripts and toxicopathic changes in Nile tilapia,Oreochromis niloticus[J].Environ Sci Pollut Res,2014,23(2):1-7.
[4]Annabi A,Said K,Messaoudi I.Heavy metal levels in gonad and liver tissues-effects on the reproductive parameters of natural populations of Aphanius facsiatus[J].Environ Sci Pollut Res,2013,20(10):7309-7319.
[5]Gill T S,Tewari H,Pande J.In vivo and in vitro effects of cadmium on selected enzymes in different organs of the fish Barbus conchonius ham(Rosy Barb)[J].Comp Biochem Physiol C,1991,100(3):501-505.
[6]Xie H,Wang J,Jiang L,et al.ROS-dependent HMGA2 upregulation mediates Cd-induced proliferation in MRC-5 cells[J].Toxicol In Vitro,2016,34:146-152.
[7]Lushchak V I.Environmentally induced oxidative stress in aquatic animals[J].Aquat Toxicol,2011,101(1):13-30.
[8]Jin Y,Liu Z,Fang L,et al.Embryonic exposure to cadmium(II)and chromium(VI)induce behavioral alterations,oxidative stress and immunotoxicity in zebrafish(Danio rerio)[J].Neurotoxicol Teratol,2015,48:9-17.
[9]Qu R,Wang X,Wang Z,et al.Metal accumulation and antioxidant defenses in the freshwater fish Carassius auratus,in response to single and combined exposure to cadmium and hydroxylated multi-walled carbon nanotubes[J].J Hazard Mater,2014,275(2):89-98.
[10]JancsóZ,Hermesz E.Impact of acute arsenic and cadmium exposure on the expression of two haeme oxygenase genes and other antioxidant markers in common carp(Cyprinus carpio)[J].J Appl Toxicol,2014,35(3):310-318.
[11]Nunes B,Caldeira C,Pereira J L,et al.Perturbations in ROS-related processes of the fish Gambusia holbrooki after acute and chronic exposures to the metals copper and cadmium[J].Environ Sci Pollut Res,2015,22(5):3756-3765.
[12]马丹旦,雷雯雯,吴昊,等.急性镉染毒对河南华溪蟹精子质量的影响[J].环境科学学报,2013,33(7):2044-2049.
[13]Wu S M,Tsai P J,Chou M Y,et al.Effects of maternal cadmium exposure on female reproductive functions,gamete quality,and offspring development in zebrafish(Danio rerio)[J].Arch Environ Contam Toxicol,2013,65(3):521-536.
[14]Simon O,Floc'H E,Geffroy B,et al.Exploring ecotoxicological fish bioassay for the evaluation of uranium reprotoxicity[J].Environ Toxicol Chem,2014,33(8):1817-1824.
[15]刘燕.水体中镉暴露对稀有鮈鲫氧化应激及组织学影响[D].重庆:西南大学,2014:17-25.
[16]Adeogun A O,Onibonoje K,Ibor O R,et al.Endocrine-disruptor molecular responses,occurrence of intersex and gonado-histopathological changes in tilapia species from a tropical freshwater dam(Awba Dam)in Ibadan,Nigeria[J].Aquat Toxicol,2016,174:10-21.
[17]Chouchene L,Banni M,Kerkeni A,et al.Cadmiuminduced ovarian pathophysiology is mediated by change in gene expression pattern of zinc transporters in zebrafish(Danio rerio)[J].Chem Biol Interact,2011,193(2):172-179.
[18]Wang Y X,Ferrari M C O,Hoover Z,et al.The effects of chronic exposure to environmentally relevant levels of waterborne cadmium on reproductive capacity and behaviour in fathead minnows[J].Arch Environ Contam Toxicol,2014,67(2):181-191.
[19]Young I S,Woodside J V.Antioxidants in health and disease[M].Lambert Academic Publishing,2013:176-186.
[20]Kusch R C,Krone P H,Chivers D P.Chronic exposure to low concentrations of waterborne cadmium during embryonic and larval development results in the long-term hindrance of antipredator behavior in zebrafish[J].Environ Toxicol Chem,2008,27(3):705-710.
[21]Vaziri N D,Lin C Y,Farmand F,et al.Superoxide dismutase,catalase,glutathione peroxidase and NADPH oxidase in lead-induced hypertension[J].Kidney Int,2003,63(1):186-194.
[22]Roméo M,Bennani N,Gnassia-Barelli M,et al.Cadmium and copper display different responses towards oxidative stress in the kidney of the sea bass Dicentrarchus labrax[J].Aquat Toxicol,2000,48(2-3):185-194.
[23]Banni M,Chouchene L,Said K,et al.Mechanisms underlying the protective effect of zinc and selenium against cadmium-induced oxidative stress in zebrafish Danio rerio[J].Biometals,2011,24(6):981-992.
[24]李文英,刘荣,蒋园,等.DBP对斑马鱼肝脏和鳃SOD及ATPase酶活性的影响[J].水生态学杂志,2007,27(4):15-18.
[25]熊道文,方涛,陈旭东,等.纳米材料对斑马鱼的氧化损伤及应激效应研究[J].环境科学,2010,31(5):1320-1327.
[26]闫博,王兰,李涌泉,等.镉对长江华溪蟹肝胰腺抗氧化酶活力的影响[J].动物学报,2007,53(6):1121-1128.
[27]马京津,徐团,轩瑞晶,等.铅对河南华溪蟹卵巢抗氧化酶活力和脂质过氧化水平的影响[J].环境科学学报,2012,32(9):2346-2352.
[28]徐团,李颖君,金芬芬.镉对长江华溪蟹精巢抗氧化酶活性及脂质过氧化的影响[J].四川动物,2011,30(5):762-764.
[29]刘娜,闫博,李涌泉,等.Cd2+对长江华溪蟹谷胱甘肽系统的影响[J].环境科学,2008,29(8):2302-2307.
[30]Iszard M B,Liu J,Klaassen C D.Effect of several metallothionein inducers on oxidative stress defense mechanisms in rats[J].Toxicology,1995,104(1-3):25-33.
[31]Zheng J L,Yuan S S,Wu C W,et al.Acute exposure to waterborne cadmium induced oxidative stress and immunotoxicity in the brain,ovary and liver of zebrafish(Danio rerio)[J].Aquat Toxicol,2016,180:36-44.
[32]Su M W,Pei R T,Yan C J.Maternal cadmium exposure induces mt2,and smt B,m RNA expression in zebrafish(Danio rerio)females and their offspring[J].Comp Biochem Physiol Cy,2012,156(156):1-6.
[33]Sellin M K,Kolok A S.Cadmium exposures during early development:Do they lead to reproductive impairment in fathead minnows[J]?Environ Toxicol Chem,2006b,25(11):2957-2963.
[34]Bourrachot S,Brion F,Pereira S,et al.Effects of depleted uranium on the reproductive success and F1generation survival of zebrafish(Danio rerio)[J].Aquat Toxicol,2014,154(1):1-11.
[35]Sellin M K,Eidem T M,Kolok A S.Cadmium exposures in fathead minnows:are there sex-specific differences in mortality,reproductive success,and Cd accumulation[J]?Arch Environ Contam Toxicol,2007,52(4):535-540.
[36]Sellin M K,Kolok A S.Cd exposures in fathead minnows:effects on adult spawning success and reproductive physiology[J].Arch Environ Contam Toxicol,2006a,51(4):594-599.
[37]Shi X,Du Y,Lam P K S,et al.Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS[J].Toxicol Appl Pharmacol,2008,230(1):23-32.
[38]鲁疆,王占洋,袁玉婷,等.氯化镉对斑马鱼胚胎的发育毒性[J].生态毒理学报,2013,8(3):381-388.
[39]Zhu B,Lei L,Li D L,et al.Developmental toxicity in rare minnow(Gobiocypris rarus)embryos exposed to Cu,Zn and Cd[J].Ecotoxicol Environ Safety,2014,104C(1):269-277.