睾酮对雄性精子发生障碍大鼠CMTM家族表达的影响

Effect of testosterone on the expression of CMTM family of the male spermatogenesis suppression rats

研究睾酮对生精功能障碍大鼠的作用,以及对CMTM家族各蛋白表达的影响。采用棉酚(50mg·kg-1)和环磷酰胺(20mg·kg-1)建立大鼠精子发生障碍模型,给予丙酸睾酮(5mg·kg-1,隔日给药,共6次)后取左侧附睾尾,收集精子。采用CASA图像分析系统对精子进行数量和活力分析,取睾丸组织采用HE染色进行形态学检查,采用蛋白印记方法(Western blotting)对CMTM家族各成员进行蛋白水平表达研究。研究发现,棉酚和环磷酰胺均可使睾丸组织发生明显的病理改变,降低精子数量和活力,而睾酮(5mg·kg-1)具有一定的改善作用。研究CMTM家族表达情况发现,棉酚和环磷酰胺均可降低雄性大鼠睾丸组织中CMTM4的表达,睾酮(5mg·kg-1)可显著提高CMTM4表达;棉酚可显著增加睾丸中CMTM2的表达,给予睾酮后CMTM2表达量显著下降,而环磷酰胺对于CMTM2影响不显著;棉酚和环磷酰胺对于CKLF1、CMTM3和CMTM5的表达影响不显著,睾丸组织中未检测到CMTM6、CMTM7和CMTM8的表达。研究结果表明,睾酮(5mg·kg-1)的生精作用可能与影响CMTM家族个别成员表达有关,趋化因子CMTM2和CMTM4可能在精子发生过程具有作用。

This study is to investigate the influence and the expression of CMTM family of testosterone on spermatogenesis suppression in the male rats treated by gossypol and cyclophosphamide. Gossypol （50 mg·kg-1） and cyclophosphamide （20 mg·kg-1） were administered to male rats to induce spermatogenesis suppression. Testosterone propionate was administrated at the dose of 5 mg·kg-1 every other day for 6 times. Sperm was collected from the left caudal epididymis,the count and motility of sperm were analyzed by CASA. Morphological change of testis tissue was observed with HE staining. The expression of CMTM family was examined by Western blotting assay. Gossypol （50 mg·kg-1） and cyclophosphamide （20 mg·kg-1） decreased the count and motility of sperm,and the pathological change of testis tissue was also observed. But,testosterone （5 mg·kg-1） had positive effect. Furthermore,CMTM4 down-expressed remarkably in the gossypol and cyclophosphamide treated rats,the expression of the CMTM4 was up-expressed after testosterone administration. On the contrary,the expression of CMTM2 increased significantly only in gossypol treated male rats,but not in cyclophosphamide treated male rats. The expression of CMTM2 was down-expressed after testosterone administration. However,no obvious change of CMTM2 was observed in cyclophosphamide treated rats. Testosterone did not influence the expression of CKLF1,CMTM3 and CMTM5,the CMTM6,CMTM7 and CMTM8 of CMTM family were not detected in testis tissue. These demonstrated that the spermatogenesis effect of testosterone （5 mg·kg-1） was associated with the expression of CMTM family,and CMTM2 and CMTM4 may take part in the spermatogenesis process.