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目的研究二硫化碳(CS2)染毒对大鼠睾丸生殖细胞凋亡线粒体途径相关基因表达的影响,探讨CS2染毒致雄性大鼠生殖系统功能紊乱的分子生物学机制。方法选取24只清洁级雄性SD大鼠,按体质量随机分为4组,即1个对照组和3个不同浓度CS2染毒组(50、250、1 250mg/m3),每日静式吸入CS22h,5d/周,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RNA,应用荧光实时定量聚合酶链反应(qPCR)检测细胞色素C(Cyto C)、半胱氨酸蛋白酶-9(Caspase-9)、半胱氨酸蛋白酶-3(Caspase-3)、凋亡诱导因子(Aif)、线粒体衍生的第2个半胱氨酸蛋白酶激活剂(Smac)、B细胞淋巴瘤/白血病-2(Bcl-2)及Bcl-2相关X基因(Bax)mRNA表达水平。结果与对照组比较,各组染毒动物睾丸生殖细胞中Caspase-9及Bax mRNA表达水平明显上升(P<0.05),Bax与Bcl-2mRNA表达水平之比亦有明显升高(P<0.05),而Smac mRNA的表达没有明显变化(P>0.05);在高浓度(1 250mg/m3)的CS2作用下,细胞中Cyto C、Caspase-3和AiF mRNA相对表达水平明显上升,而Bcl-2mRNA表达水平则明显下降,差异均有统计学意义(P<0.05)。结论 CS2染毒可影响细胞凋亡线粒体途径相关基因的转录水平,这可能是CS2所致雄性生殖系统毒性的作用机制之一。
Objective To investigate the effects of CS2 exposure on the expression of mitochondria-related genes in germ cells of testis germ cells in rats and to explore the molecular mechanism of reproductive system dysfunction induced by CS2 in rats. Methods Twenty-four clean male Sprague-Dawley rats were randomly divided into four groups according to body weight: one control group and three CS2-treated groups (50, 250 and 250 mg / m3) CS22h, 5d / week, co-administration of 10 weeks; control group only inhaled air. Rats were killed after decapitation by decapitation, and testicular tissue was taken for RNA extraction. Cyto C, Caspase-9, Caspase-3, Aif, mitochondria-derived second cysteine proteinase activator (Smac), B cell lymphoma / leukemia-2 (Bcl-2 ) And Bcl-2 related gene X (Bax) mRNA expression levels. Results Compared with the control group, the expression of Caspase-9 and Bax mRNA in testis germ cells in each group increased significantly (P <0.05) and the ratio of Bax to Bcl-2 mRNA expression increased significantly (P <0.05) , While the expression of Smac mRNA did not change significantly (P> 0.05). The relative expression of Cyto C, Caspase-3 and AiF mRNA increased significantly in CS2 cells exposed to high concentration of 1,250 mg / m3, while the expression of Bcl-2 mRNA The expression level was significantly decreased, the differences were statistically significant (P <0.05). Conclusion CS2 can affect the transcription level of mitochondrial pathway-related genes in apoptosis, which may be one of the mechanisms of CS2-induced toxicity of male reproductive system.