高效氯氰菊酯对小鼠卵巢生殖功能的影响及维生素E的干预作用
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摘要
为探究高效氯氰菊酯(beta-cypermethrin,β-CP)对雌性小鼠卵巢生殖功能的影响及维生素E (vitamin E, VE)的干预作用,将雌性昆白小鼠随机分为6组:空白对照组(花生油处理)、β-CP不同剂量(10、20、40 mg/kg)处理组、VE保护组(20 mg/kgβ-CP+20 mg/kg VE)和VE组(20 mg/kg VE),连续灌胃10 d。灌胃结束后取小鼠卵巢组织,观察组织结构的病理变化,采用免疫组化法、蛋白免疫印迹试验及RT-PCR方法检测卵巢中StAR蛋白含量及casp-3、casp-8、INHα和INHβB基因mRNA表达的变化。结果显示:与对照相比,10、20和40 mg/kg的β-CP处理均使小鼠卵巢组织结构发生了损伤,使组织中StAR蛋白的浓度分别降低了18.8%、36.3%和40.3%,casp-3基因的表达分别升高了16.0%、26.7%和52.9%,INHα基因的表达分别升高了34.5%、83.6%和228.7%,INHβB基因的表达分别升高了7.5%、39.2%和52.7%;20和40 mg/kg的β-CP处理使得casp-8基因的表达分别升高了27.1%和36.7%。上述处理组与对照组的差异均达显著水平(P<0.05)。与20 mg/kgβ-CP处理组相比,VE保护组的StAR蛋白含量也显著增多(P<0.05)。研究表明,β-CP对小鼠卵巢具有毒性作用,这与β-CP抑制StAR蛋白的合成,上调casp-3、casp-8、INHα及INHβB基因的表达有关;添加VE对小鼠卵巢有一定的保护作用,这与VE可减弱β-CP对StAR蛋白合成的抑制作用有关。
The study was conducted to investigate the influence of beta-cypermethrin(β-CP) on ovary reproductive function and the intervention of vitamin E(VE) in mice. Blank control(peanut oil treatment), three different doses of β-CP(10, 20 and 40 mg/kg), protection(20 mg/kg β-CP+20 mg/kg vitamin E) and vitamin E(20 mg/kg) were applied to 6 groups of mice for 10 d continuously. After the treatment, the ovarian tissue was obsoleted to observe the pathological changes of ovarian tissue structure. The changes of StAR protein content, casp-3, casp-8, INHα and INHβB gene expression were determined by immunocytochemistry, Western blotting and RT-PCR. The results showed that,compared with the control group, the treatment of β-CP at 10, 20 and 40 mg/kg damaged the ovarian tissue in mice, the concentration of StAR protein was reduced by 18.8%, 36.3% and 40.3%, the expression of casp-3 was increased by 16.0%, 26.7% and 52.9%, the expression of INHα was increased by 34.5%, 83.6% and 228.7%, the expression of INHβB was increased by 7.5%, 39.2% and 52.7%; the treatment of β-CP at 20 and 40 mg/kg increased the expression of casp-8 by 27.1% and 36.7%. There are significant differences between the above treatment groups and control groups(P < 0.05). The concentrarion of StAR protein increased in the protection group compared with the 20 mg/kg β-CP treatment group(P < 0.05). Those results demonstrated that β-CP had a toxic effect on the ovarian tissue of mice, which was caused by the inhibition of StAR protein synthesis by β-CP and the increase in the expression of casp-3, casp-8, INHα and INHβB. Vitamin E had a protection effect on mice ovarian tissue after, which resulted from the decrease of the inhibit effect of β-CP on StAR protein by vitamin E.
The study was conducted to investigate the influence of beta-cypermethrin(β-CP) on ovary reproductive function and the intervention of vitamin E(VE) in mice. Blank control(peanut oil treatment), three different doses of β-CP(10, 20 and 40 mg/kg), protection(20 mg/kg β-CP+20 mg/kg vitamin E) and vitamin E(20 mg/kg) were applied to 6 groups of mice for 10 d continuously. After the treatment, the ovarian tissue was obsoleted to observe the pathological changes of ovarian tissue structure. The changes of StAR protein content, casp-3, casp-8, INHα and INHβB gene expression were determined by immunocytochemistry, Western blotting and RT-PCR. The results showed that,compared with the control group, the treatment of β-CP at 10, 20 and 40 mg/kg damaged the ovarian tissue in mice, the concentration of StAR protein was reduced by 18.8%, 36.3% and 40.3%, the expression of casp-3 was increased by 16.0%, 26.7% and 52.9%, the expression of INHα was increased by 34.5%, 83.6% and 228.7%, the expression of INHβB was increased by 7.5%, 39.2% and 52.7%; the treatment of β-CP at 20 and 40 mg/kg increased the expression of casp-8 by 27.1% and 36.7%. There are significant differences between the above treatment groups and control groups(P < 0.05). The concentrarion of StAR protein increased in the protection group compared with the 20 mg/kg β-CP treatment group(P < 0.05). Those results demonstrated that β-CP had a toxic effect on the ovarian tissue of mice, which was caused by the inhibition of StAR protein synthesis by β-CP and the increase in the expression of casp-3, casp-8, INHα and INHβB. Vitamin E had a protection effect on mice ovarian tissue after, which resulted from the decrease of the inhibit effect of β-CP on StAR protein by vitamin E.
引文
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[21]HUANG F,LIU Q Y,XIE S J,et al.Cypermethrin induces macrophages death through cell cycle arrest and oxidative stressmediated JNK/ERK signaling regulated apoptosis[J].Int J Mol Sci,2016,17(6):885.
[22]WANG X Z,LIU S S,SUN Y,et al.Beta-Cypermethrin impairs reproductive function in male mice by inducing oxidative stress[J].Theriogenology,2009,72(5):599-611.
[23]HOCINE L,MERZOUK H,MERZOUK S A,et al.The effects of alpha-cypermethrin exposure on biochemical and redox parameters in pregnant rats and their newborns[J].Pestic Biochem Physiol,2016,134:49-54.
[24]GIRAY B,GüRBAY A,HINCAL F.Cypermethrin-induced oxidative stress in rat brain and liver is prevented by vitamin E or allopurinol[J].Toxicol Lett,2001,118(3):139-146.
[25]PETERSON G L.A simplification of the protein assay method of Lowry,et al which is more generally applicable[J].Anal Biochem,1977,83(2):346-356.
[26]ZHOU Y J,WANG X D,XIAO S,et al.Exposure to betacypermethrin impairs the reproductive function of female mice[J].Regul Toxicol Pharmacol,2018,95:385-394.
[27]ZHOU Y J,WANG J H,WANG L Q,et al.Effect of betacypermethrin exposure on embryo implantation in mice[J].Reprod Toxicol,2018,76:1-11.
[28]SOCCIO R E,BRESLOW J L.Intracellular cholesterol transport[J].Arterioscler Thromb Vasc Biol,2004,24(7):1150-1160.
[29]TSUCHIYA M,INOUE K,MATSUDA H,et al.Expression of steroidogenic acute regulatory protein(StAR)and LH receptor in MA-10 cells[J].Life Sci,2003,73(22):2855-2863.
[30]MARETTOVA E,MARETTA M,LEGáTH J.Effect of pyrethroids on female genital system[J].Anim Reprod Sci,2017,184:132-138.
[31]DE JONG F H.Inhibin[J].Physiol Rev,1988,68(2):555-607.
[2]JIN T,LIN Y Y,JIN Q A,et al.Population susceptibility to insecticides and the development of resistance in Bactrocera cucurbitae(Diptera:Tephritidae)[J].J Econ Entomol,2016,109(2):837-846.
[3]FRANCO A A,ZANARDI O Z,JACOB C R O,et al.Susceptibility of Euseius concordis(Mesostigmata:Phytoseiidae)to pesticides used in citrus production systems[J].Exp Appl Acarol,2017,73(1):61-77.
[4]QU Y Y,XIAO D,LIU J J,et al.Sublethal and hormesis effects of beta-cypermethrin on the biology,life table parameters and reproductive potential of soybean aphid Aphis glycines[J].Ecotoxicology,2017,26(7):1002-1009.
[5]LENG G,BERGER-PREIβE,LEVSEN K,et al.Pyrethroids used indoor-ambient monitoring of pyrethroids following a pest control operation[J].Int J Hyg Environ Health,2005,208(3):193-199.
[6]SCOTT J G,YOSHIMIZU M H,KASAI S.Pyrethroid resistance in Culex pipiens mosquitoes[J].Pestic Biochem Physiol,2015,120:68-76.
[7]WANG X,HE B N,KONG B D,et al.β-Cypermethrin and its metabolite 3-phenoxybenzoic acid exhibit immunotoxicity in murine macrophages[J].Acta Biochimica Biophysica Sinica(Shanghai),2017,49(12):1083-1091.
[8]HUGHES M F,ROSS D G,STARR J M,et al.Environmentally relevant pyrethroid mixtures:a study on the correlation of blood and brain concentrations of a mixture of pyrethroid insecticides to motor activity in the rat[J].Toxicology,2016,359-360:19-28.
[9]MAURYA S K,MISHRA J,ABBAS S,et al.Cypermethrin stimulates GSK3β-dependent Aβand p-tau proteins and cognitive loss in young rats:reduced HB-EGF signaling and downstream neuroinflammation as critical regulators[J].Mol Neurobiol,2016,53(2):968-982.
[10]BASKAR M K,MURTHY P B.Acute in vitro neurotoxicity of some pyrethroids using microelectrode arrays[J].ToXicology in Vitro,2018,47:165-177.
[11]AL-HAMDANI N M H,NARASINHACHARY Y H.Endocrine disruptive action of cypermethrin in male mice[J].Toxicol Environ Health Sci,2011,3(2):69-79.
[12]CALDERóN-SEGURA M E,GóMEZ-ARROYO S,CORTéS-ESLAVA J,et al.In vitro cytotoxicity and genotoxicity of Furia?180SC(zeta-cypermethrin)and Bulldock 125?SC(β-cyfluthrin)pyrethroid insecticides in human peripheral blood lymphocytes[J].Toxicol Mech Methods,2018,28(4):268-278.
[13]HUSSIEN H M,ABDOU H M,YOUSEF M I.Cypermethrin induced damage in genomic DNA and histopathological changes in brain and haematotoxicity in rats:the protective effect of sesame oil[J].Brain Res Bull,2013,92:76-83.
[14]SEIFERTOVáM,?ECHOVáE,LLANSOLA M,et al.Determination of selected neurotoxic insecticides in small amounts of animal tissue utilizing a newly constructed mini-extractor[J].Anal Bioanal Chem,2017,409(25):6015-6026.
[15]YUN X M,HUANG Q C,RAO W B,et al.A comparative assessment of cytotoxicity of commonly used agricultural insecticides to human and insect cells[J].Ecotoxicol Environ Saf,2017,137:179-185.
[16]WANG Q,XU L F,ZHOU J L,et al.Antagonism effects of cypermethrin on interleukin-6-induced androgen receptor activation[J].Environ Toxicol Pharmacol,2015,40(1):172-174.
[17]MANNA S,BHATTACHARYYA D,MANDAL T K,et al.Repeated dose toxicity of alfa-cypermethrin in rats[J].J Vet Sci,2004,5(3):241-245.
[18]WEI K Q,YANG J X.Oxidative damage induced by copper and beta-cypermethrin in gill of the freshwater crayfish Procambarus clarkii[J].Ecotoxicol Environ Saf,2015,113:446-453.
[19]ZHANG J Y,LIU L L,REN L,et al.The single and joint toxicity effects of chlorpyrifos and beta-cypermethrin in zebrafish(Danio rerio)early life stages[J].J Hazard Mater,2017,334:121-131.
[20]CHEN L,XU P,DIAO J L,et al.Distribution,metabolism and toxic effects of beta-cypermethrin in lizards(Eremias argus)following oral administration[J].J Hazard Mater,2016,306:87-94.
[21]HUANG F,LIU Q Y,XIE S J,et al.Cypermethrin induces macrophages death through cell cycle arrest and oxidative stressmediated JNK/ERK signaling regulated apoptosis[J].Int J Mol Sci,2016,17(6):885.
[22]WANG X Z,LIU S S,SUN Y,et al.Beta-Cypermethrin impairs reproductive function in male mice by inducing oxidative stress[J].Theriogenology,2009,72(5):599-611.
[23]HOCINE L,MERZOUK H,MERZOUK S A,et al.The effects of alpha-cypermethrin exposure on biochemical and redox parameters in pregnant rats and their newborns[J].Pestic Biochem Physiol,2016,134:49-54.
[24]GIRAY B,GüRBAY A,HINCAL F.Cypermethrin-induced oxidative stress in rat brain and liver is prevented by vitamin E or allopurinol[J].Toxicol Lett,2001,118(3):139-146.
[25]PETERSON G L.A simplification of the protein assay method of Lowry,et al which is more generally applicable[J].Anal Biochem,1977,83(2):346-356.
[26]ZHOU Y J,WANG X D,XIAO S,et al.Exposure to betacypermethrin impairs the reproductive function of female mice[J].Regul Toxicol Pharmacol,2018,95:385-394.
[27]ZHOU Y J,WANG J H,WANG L Q,et al.Effect of betacypermethrin exposure on embryo implantation in mice[J].Reprod Toxicol,2018,76:1-11.
[28]SOCCIO R E,BRESLOW J L.Intracellular cholesterol transport[J].Arterioscler Thromb Vasc Biol,2004,24(7):1150-1160.
[29]TSUCHIYA M,INOUE K,MATSUDA H,et al.Expression of steroidogenic acute regulatory protein(StAR)and LH receptor in MA-10 cells[J].Life Sci,2003,73(22):2855-2863.
[30]MARETTOVA E,MARETTA M,LEGáTH J.Effect of pyrethroids on female genital system[J].Anim Reprod Sci,2017,184:132-138.
[31]DE JONG F H.Inhibin[J].Physiol Rev,1988,68(2):555-607.