二硫化碳对大鼠睾丸生殖细胞凋亡中线粒体途径相关基因表达的影响  

Effect of carbon disulfide on the expression of mitochondrion related gene in mediating testicular germ cells apoptosis of male rats

作　　者：黄晓彧[1] 张振[1] 王为[1] 董煜[1] 郭寅生[1] 彭小波[1] 吴卓泽 许忠宇[1] 彭钊[1] 陈国元[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院,湖北武汉430030

出　　处：《工业卫生与职业病》2014年第2期108-113,共6页Industrial Health and Occupational Diseases

基　　金：国家自然科学基金资助项目(30872091;81172637);华中科技大学大学生创新创业项目基金资助项目(12B411)

摘　　要：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 effect of carbon disulfide on the expression of mitochondrion re- lated genes in mediating testicular germ cell apoptosis of male rats, and molecular biological mechanism of re- productive toxicity induced by CS2 in male rats. Methods Twenty four SD male rats were divided into three CS2 exposed groups and a control group. Rats were exposed to multiple levels of CS2 concentration（0,50,250, and 1250 mg/m3） by inhalation for 10 weeks, 5d/w, 2h/d. After exposure, rats were sacrificed by decapita- tion, RNA in testis were extracted. The expressions of several genes, such as Cyto C, Caspase--9, Caspase --3, AIF, Smac, Bcl--2 and Bax, were detected by real--time fluorescent quantitative polymerase chain re- action （qPCR）. Results Compared with the controls, the levels of Caspase--9 and Bax mRNA in testicular germ cells, as well as the ratio between Bax and Bcl--2, increased when rats were exposed to different doses of CS2 （P〈0.05）, while the levels of Smac mRNA showed no statistical differences （P〉0.05）. Related ex- pression levels of Cyto C, Caspase--3 and Air mRNA in testicular germ cells increased, while those of Bcl-- 2 mRNA significantly decreased, as rats were exposed to the high dose of CS2 （1250 mg/m3 ）. Conclusions CS2exposure could affect the expression of genes related to mitochondrial mediated apoptosis, and accordingly result in mitochondrial mediated apoptosis in testicular germ cells of male rats.

关 键 词：二硫化碳 生殖细胞 线粒体 细胞凋亡 基因 

分 类 号：R114[医药卫生—卫生毒理学]