畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (4): 500-504.doi:

• 基础兽医 • 上一篇    下一篇

线粒体凋亡途径在锰致鸡支持-生精细胞凋亡中的作用

毕明玉,李金龙,李术,陈蕾,张子威,唐洪鹏,徐世文*   

  1. 东北农业大学动物医学学院,哈尔滨 150030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-04-25 发布日期:2010-04-25
  • 通讯作者: 徐世文

The Role of Mitochondria Mediated Apoptosis Pathway in Sertoli-germ CellApoptosis Induced by Manganese in Cocks

BI Ming-yu, LI Jin-long, LI Shu, CHEN Lei, ZHANG Zi-wei, TANG Hong-peng, XU Shi-wen*   

  1. College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-04-25 Published:2010-04-25

摘要: 探讨线粒体凋亡途径在锰致鸡支持-生精细胞凋亡中的作用。取对数生长期鸡支持-生精细胞,用含0、2、3、4 mmol·L-1 MnCl2的DMEM培养液,培养24 h,采用吖啶橙-溴化乙锭(AO/EB)双荧光染色法检测鸡支持-生精细胞凋亡,流式细胞仪检测线粒体膜电位(△ψm),Western blot法检测胞浆内细胞色素C(Cytc)蛋白表达含量,分光光度法检测半胱氨酸天冬氨酸蛋白酶9,3(Caspase-9,3)活性。用含MnCl2的DMEM培养液培养24 h后,处理组细胞与对照组相比,鸡支持-生精细胞凋亡指数逐渐升高(P<0.01);线粒体膜电位显著降低(P<0.01);胞浆中Cytc蛋白表达量逐渐增加;Caspase-9,3活性逐渐升高(P<0.01)。线粒体途径介导的凋亡是锰对鸡生殖毒性作用的主要机制之一。

Abstract: The aim of this study was to discuss the role of mitochondria mediated apoptosis pathway in Sertoli-germ cell apoptosis induced by manganese in cocks. Cock Sertoli-germ cells were cultivated in the DMEM for 24 h added with MnCl2, whose final concentrations were 0, 2, 3, and 4 mmol·L-1 respectively. The apoptosis was detected by AO/EB double staining. Mitochondrial transmembrane potential was detected by flow cytometer. Expression of Cytochrome c (Cytc) in cytolymph was detected by western blotting. The activities of Caspase-9, 3 were examined by spectrophotography. Compared with control group, the apoptosis index (AI) of cock Sertoli-germ cells was significantly increased (P<0.01), mitochondrial transmembrane potential was significantly decreased (P<0.01), expression of Cytc in cytolymph was increased, and the activity of Caspase-9, 3 were increased (P<0.01) in 2, 3, and 4 mmol·L-1 MnCl2 group. The apoptosis mediated by mitochondria pathway was one of the reproductive toxic mechanisms caused by manganese.