CAJ | 학술논문

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) MnCl_2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一生精细胞凋亡,流式细胞仪检测线粒体膜电位(△_(ψ_m)),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MnCl_2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.
탐토선립체조망도경재맹치계지지-생정세포조망중적작용.취대수생장기계지지-생정세포,용함0、2、3、4 mmol·L~(-1) MnCl_2적DMEM배양액,배양24 h,채용아정등-추화을정(AO/EB)쌍형광염색법검측계지지일생정세포조망,류식세포의검측선립체막전위(△_(ψ_m)),Western blot법검측포장내세포색소C(Cytc)단백표체함량,분광광도법검측반광안산천동안산단백매9,3(Caspase-9,3)활성.용함MnCl_2적DMEM배양액배양24 h후,처리조세포여대조조상비,계지지-생정세포조망지수축점승고(P<0.01);선립체막전위현저강저(P<0.01);포장중Cytc단백표체량축점증가;Caspase-9,3활성축점승고(P<0.01).선립체도경개도적조망시맹대계생식독성작용적주요궤제지일.
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 MnCl_2,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) MnCl_2 group.The apoptosis mediated by mitochondria pathway was one of the reproductive toxic mechanisms caused by manganese.