阿特拉津对大鼠支持细胞凋亡的影响  

Effect of atrazine on apoptosis in cultured rat Sertoli cells

作　　者：宋阳[1] 张誉[1] 马梦婷 张立实[1] Song Yang Zhang Yu Ma Mengting Zhang Lishi(Siehuan Center for Disease Control and Prevention, Chengdu 610041 , China)

机构地区：[1]四川省疾病预防控制中心,成都610041

出　　处：《卫生研究》2017年第1期103-108,共6页Journal of Hygiene Research

基　　金：国家自然科学基金重点项目(No.81030053)

摘　　要：目的探讨阿特拉津对雄性SD大鼠睾丸支持细胞凋亡的影响及其作用机制。方法选用18~21日龄雄性SD大鼠,建立支持细胞原代培养模型。设溶剂对照组和阿特拉津10、50、100和200μmol/L 4个染毒剂量组,对支持细胞染毒24 h后,采用MTT法检测细胞活性,免疫荧光法观察波形蛋白的表达,流式细胞术测定细胞凋亡率,Real time-PCR与ELISA分别检测Fas L、caspase-3与caspase-8的mRNA表达及蛋白含量。结果阿特拉津50、100和200μmol/L染毒组细胞活力分别为82.47%、75.23%和53.76%,与溶剂对照组比较分别降低17.53%(P<0.05)、24.77%(P<0.05)、46.24%(P<0.01)。波形蛋白表达受抑制。阿特拉津50、100和200μmol/L染毒组细胞凋亡率分别为8.53%、13.07%和14.45%,与溶剂对照组比较分别升高86.86%(P<0.01)、186.13%(P<0.01)、216.06%(P<0.01)。Fas L、caspase-3、caspase-8的mRNA表达及蛋白含量上调(P<0.05或P<0.01)。结论阿特拉津可影响大鼠支持细胞活性,可通过Fas L途径诱导支持细胞凋亡。Objective To elucidate the effects of atrazine（ ATZ） on apoptosis in cultured rat Sertoli cells. Methods The Sertoli cells isolated from male SD rats（ 18-21day） were used to set up Sertoli cells primary culture model in vitro,and these cells were treated with 0,10,50,100 and 200 μmol / L ATZ for 24 h. Cell viability was assessed by MTT assay,and the vimentin expression of Sertoli cells was analyzed by immunofluorescence assay,and apoptosis were evaluated by flow cytometry. Real timePCR was used to analyze the mRNA levels of Fas L,caspase-3 and caspase-8,and the protein contents of Fas L,caspase-3 and caspase-8 were determined by ELISA. Results The viability of Sertoli cells of 50,100 and 200 μmol / L ATZ groups were 82. 47%,75. 23% and 53. 76%. Compared with the control group,the viability of Sertoli cells decreased by 17. 53%（ P〈0. 05）,24. 77%（ P〈0. 05） and 46. 24%（ P〈0. 01）,respectively. The apoptosis rates of Sertoli cell of 50,100 and 200 μmol / L ATZ groups were 8. 53%,13. 07% and 14. 45%. Compared with the control group,the apoptosisrates of Sertoli cells were increased by 86. 86%（ P〈0. 01）,186. 13%（ P〈0. 01）,216. 06%（ P〈0. 01）,respectively. the mRNA and protein expressions of Fas L,caspase-3 and caspase-8 were significantly increased at 50,100 and 200 μmol / L ATZ groups（ P〈0. 05,P〈0. 01）. The Vimentin expression of Sertoli cells was inhibited in treated groups. Conclusion ATZ can affect the viability of Sertoli cells,and induce apoptosis of Sertoli cells through a Fas L dependent pathway.

关 键 词：阿特拉津 支持细胞 波形蛋白 细胞凋亡 FASL 

分 类 号：R994.6[医药卫生—毒理学] S482.42[医药卫生—药学]