二硫化碳对大鼠睾丸组织胞浆线粒体凋亡相关蛋白的影响

Effect of carbon disulfide on mitochondrial apoptosis-related proteins in cytoplasm of testicular tissue among male rats

目的 探讨线粒体途径在二硫化碳（CS2）吸入染毒致大鼠睾丸生殖细胞凋亡中的作用.方法 选取24只清洁级雄性SD大鼠按体重随机分为4组,每组6只,即设3个不同浓度CS2染毒组（50、250、1250 mg/m3）和1个对照组,每日静式吸入CS22 h,每周5d,共染毒10周,对照组仅吸入空气.染毒结束后断头处死大鼠,取睾丸组织,提取组织胞浆蛋白,用蛋白印迹分析检测AIF、Cyto C、Bcl-2、Bax、Procaspase-9和Procaspase-3含量,并用试剂盒检测Caspase-9和Caspase-3活力.结果 与对照组相比,各染毒组动物睾丸组织胞浆中Cyto C含量有明显上升,差异有统计学意义（P＜0.05）；在中浓度组（250 mg/m3）中,Bax/Bcl-2比值、Caspase-9和Caspase-3活力上升,差异有统计学意义（P＜0.05）,而Procaspase-9和Procaspse-3蛋白含量下降,差异有统计学意义（P＜0.05）；在高浓度（1250 mg/m3）的CS2作用下,胞浆中Bax和AIF蛋白的相对表达水平明显上升,而Bcl-2蛋白表达水平则呈明显下降,这些变化的差异均有统计学意义（P＜0.05）.结论 线粒体途径在CS2所致大鼠睾丸组织细胞凋亡中发挥着重要作用.

Objective To investigate the role of mitochondrial pathway in the apoptosis of spermatogenic cells induced by inhalation of carbon disulfide in male rats.Methods Twenty-four male Sprague-Dawley rats （clean grade） were divided into four groups according to their body weights：three CS2 exposure groups （CS2 concentrations：50,250,and 1250 mg/m3） and a control group.The rats in CS2 exposure groups were exposed to CS2 by static inhalation for 10 weeks （2 h/d,5 d/w）,while the rats in control group were exposed to air.Then,all rats were sacrificed by decapitation; testicular tissues were collected,and cytoplasmic proteins were extracted;the levels of apoptosis-inducing factor （AIF）,cytochrome c （cyto c）,Bcl-2,Bax,procaspase-9,and procaspase3 were measured by Western blot,and the activities of caspase-9 and caspase-3 were measured using a test kit.Results Compared with the control group,all CS2 exposure groups had significantly increased levels of cyto c in the cytoplasm of testicular tissue （P＜0.05）; in the 250 mg/m3 CS2 exposure group,the Bax/Bcl-2 ratio and activities of caspase-9 and caspase-3 increased significantly （P＜0.05）,and the content of procaspase-9 and procaspase-3 decreased significantly （P＜0.05）; in the 1250 mg/m3 CS2 exposure group,the relative expression levels of Bax and AIF in cytoplasm increased significantly （P＜0.05）,and the expression level of Bcl-2 decreased significantly （P＜0.05）.Conclusion Mitochondrial pathway plays an important role in the CS2-induced apoptosis of spermatogenic cells in testicular tissue among male rats.