二硫化碳对大鼠睾丸支持-生精共培养细胞凋亡线粒体途径相关基因表达的影响

张振; 王为; 董煜; 黄晓彧; 郭寅生; 周义军; 陈国元

二硫化碳对大鼠睾丸支持-生精共培养细胞凋亡线粒体途径相关基因表达的影响

Effect of Carbon Disulfide on Expression of Genes Related with Mitochondrion Mediated Sertoli-Germ Cells Apoptosis in Male Rats

摘要

摘要:
目的研究二硫化碳(carbon disulfide, CS₂)染毒对大鼠睾丸支持-生精共培养细胞凋亡线粒体途径相关基因表达的影响。

方法选用22~24 d龄清洁级雄性SD大鼠,建立睾丸生精-支持细胞体外共培养体系,将细胞分为4组:1个对照组和3个不同浓度CS₂染毒组(1、2、4μmol/mL),提取RNA,应用荧光实时定量聚合酶链反应(RT-PCR)检测Cyto C、Apaf-1、Caspase-3、Caspase-9、Aif、EndoG、Smac、IAPS、Bax、Bcl-2 mRNA表达水平。

结果大鼠睾丸支持-生精共培养细胞中, CS₂染毒可导致Cyto C、Smac、IAPS、Caspase-3、Bax mRNA的表达水平和Bax/Bcl-2 mRNA表达水平值均明显升高(P>0.05),同时使Apaf-1、Aif、Bcl-2和EndoG mRNA的表达水平明显下降(P>0.05)。

结论 CS₂染毒可影响大鼠睾丸支持-生精共培养细胞凋亡线粒体途径相关基因的表达,并且这一机制与CS₂导致生殖损伤相关。

Abstract:
Objective To study the effects of carbon disulfide (CS₂) on the expression of genes which are related with mitochondrion mediated sertoli-germ cells apoptosis in male rats.

Methods A 22-24 day-old SD male rat of clean grade was selected to establish the rat sertoli-germ cells co-culture system in vitro. According to the designed concentrations of CS₂ (0, 1, 2, 4μmol/mL), the cells were divided into four groups accordingly and RNA were extracted. The mRNA expression levels of Cyto C, Apaf-1, Caspase-3, Caspase-9, Aif, EndoG, Smac, IAPS, Bax, Bcl-2 were detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR).

Results Comparing with the control group, the levels of Cyto C, Smac, IAPS, Caspase-3, Bax, and Bax/Bcl-2 mRNA in the sertoli-germ cells were increased, while the levels of Apaf-1, Aif, Bcl-2, and EndoG mRNA were decreased in CS₂ exposure groups (P>0.05).

Conclusion CS₂ exposure affects the expression of genes related with mitochondrial mediated apoptosis, and it probably relates to reproductive toxicity induced by CS₂.

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