雌激素及其受体ERβ在氰戊菊酯所致雄性大鼠生殖毒性中的作用  

Effects of Estrogen and ERβon Reproductive Toxicity Induced by Fenvalerate in Male Rats

作　　者：郭茂 陈琰 李兴元 孔德营 GUO Mao;CHEN Yan;LI Xingyuan;KONG Deying(Department of Physiology,Basic Medical College,Zunyi Medical University,Zunyi Guizhou 563000,China)

机构地区：[1]遵义医科大学基础医学院生理学教研室,贵州遵义563000

出　　处：《西南大学学报（自然科学版）》2021年第10期66-76,共11页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(31760339);贵州省科技厅联合基金项目(CK-1044);贵州省教育厅青年人才成长项目(CJ-910).

摘　　要：为探讨雌激素及其受体ERβ在氰戊菊酯(Fen)致雄性SD大鼠生殖毒性中的作用,采用SD大鼠将其随机分为对照组(Fen 0 mg/kg)、低剂量Fen染毒组(20 mg/kg)、高剂量Fen染毒组(40 mg/kg),每组8只,染毒组以玉米油溶解配制成不同浓度的Fen溶液等体积灌胃,对照组给予等量玉米油,每日灌胃1次,连续染毒30 d后,采用常规方法检测精子数量、精子活力及畸形率,ELISA测定雄鼠血清和睾丸中雌二醇(E2)的水平,Real-time PCR和Western Bolt检测芳香化酶(CYP19A1),ERβ,Bax,Bcl-2,Caspase 3及Caspase 9的mRNA和蛋白表达水平,TUNEL法检测大鼠睾丸生殖细胞凋亡情况,HE染色法观察睾丸病理结构.结果显示:Fen染毒30 d后,与对照组相比,在40 mg/kg组精子数量、a级精子活力及精子活动率均显著降低,精子畸形率显著增加(p<0.05);血清和睾丸E2水平均显著升高(p<0.05,p<0.01);CYP19A1 mRNA和蛋白表达水平在20 mg/kg和40 mg/kg组显著增加(p<0.05或p<0.01),ERβmRNA和蛋白表达水平在40 mg/kg组均显著增加(p<0.01);大鼠睾丸Bax,Caspase 3及Caspase 9的mRNA和蛋白表达水平在40 mg/kg组均显著增加(p<0.01),Bcl-2 mRNA和蛋白表达水平在40 mg/kg组均显著降低(p<0.01);TUNEL结果显示:大鼠睾丸生殖细胞凋亡增加且在40 mg/kg组差异有统计学意义(p<0.01);睾丸病理结构发现,染毒组大鼠睾丸曲细精管生精细胞数量减少或缺失,管腔内精子减少,部分曲细精管生精细胞排列紊乱.以上结果表明,Fen可能通过上调雄性SD大鼠睾丸CYP19A1的表达,增加雄鼠睾丸E2的水平,同时,上调雄鼠睾丸ERβ的表达,过度增强雌激素-ERβ信号作用,促进生精细胞的凋亡,引起精子发生异常.To explore the roles of estrogen and its receptor ERβ(estrogen receptorβ)in reproductive toxicity of fenvalerate in male SD rats,in this study,twenty-four male SD rats were randomly divided into three groups with eight rats in each,one control group and two treatmentgroups administered with fenvalerate at 20 and 40 mg/kg,respectively,by gavage for 30 consecutive days.Semen quality parameters were measured with conventional optical microscopy;the level of E2 in the serum and the testes was tested by enzyme-linked immunosorbent assay(ELISA);the mRNA and protein expression of CYP19A1,ERβ,Bax,Bcl-2,Caspase3 and Caspase9 were detected by real-time PCR and Western blotting;histological changes in the testes were observed by hematoxylineosin(HE)staining,and germ cell apoptosis was measured by TdT-mediated dUTP Nick-End Labeling(TUNEL).The results showed that compared with the control group,the 40 mg/kg fenvalerate group had significantly reduced sperm count and sperm motility(p<0.05),and had significantly higher levels of sperm malformation(p<0.05)and E2 levels in the serum(p<0.05)and the testicular(p<0.01).TUNEL-positive germ cells significantly increased in the 40 mg/kg fenvalerate exposure group,and the mRNA and protein expression of proapoptotic gene Bax,Caspase 3 and Caspase 9 in rat testes were also significantly up-regulated,the expression levels of antiapoptotic gene Bcl-2 was significantly reduced.Meanwhile,the number of spermatogenic cells and spermatozoa in the seminiferous tubules decreased in the fenvalerate exposure groups.These results indicate that fenvalerate can increase the level of E2 in male SD rats by up-regulating the expression of CYP19A1 and,in addition,it can also up-regulate the expression of ERβin the testis,so as to increase the combination of E2 and ERβ,promote the apoptosis of spermatogeniccells,and cause spermatogenesis disorder.

关 键 词：氰戊菊酯 雌激素 雌激素受体 生殖毒性 

分 类 号：Q492.4[生物学—生理学]