金匮肾气丸对小鼠生精恢复的基因表达谱研究

目的:研究金匮肾气丸对无精子症模型小鼠生精能力恢复作用的睾丸全基因表达谱.方法:12周龄NIH雄性小鼠按照40 mg· kg -单次ip 2％白消安药液制备无精子症小鼠动物模型,观察42 d后,给予金匮肾气丸中药1 g·kg-1,ig给药78 d(2个生精周期),以造模后模型组(自然恢复)、溶媒和正常动物为3种对照,通过基因芯片技术检测服用金匮肾气丸的无精子小鼠与正常对照小鼠之间睾丸中差异表达的基因,对这些基因按照表达差异程度、功能注释和Pathway进行分析.结果:动物实验结果显示:无精子症模型小鼠经过78 d的金匮肾气丸ig给药后,在动物体质量、生殖器官组织质量和指数、附睾精子质量(活动率、活动力、浓度)等与生殖能力相关的指标上与模型组比较有明显的改善.基因表达谱芯片结果显示:服用金匮肾气丸的无精子小鼠与正常组相比,睾丸中差异表达2倍以上的基因804个(占2.53％),其中上调517个,低于正常表达1/2的287个,差异表达4倍以上的基因134个(占0.42％)其中上调127个,低于正常表达1/4的7个,提示以正向促进/上调作用为主,以负向抑制/下调作用为辅；主要通过对细胞周期、细胞因子-细胞因子受体作用、Ecm受体作用、Jak-Stat信号通路、Mapk信号通路、Notch信号通路、Toll样受体信号通路和Wnt信号通路等8个信号通路系统的调节实现生精能力的提高.结论:金匮肾气丸有助于促进无精子小鼠生精能力的恢复.通过基因芯片检测后与正常小鼠相比,这种促进作用可能是通过多途径对生精于细胞中与细胞增殖分裂相关基因的正向调节实现的.

Gene Expression Profile of Jinkui Shenqi Wan on the Spermatogenesis Recovery in Azoospermia Mice

Objective: To study the gene expression profile of Jinkui Shenqi Wan on the recovery of spermatogenesis in azoospermia mice. Method: Liquid busulfan 2% was injected (ip 40 mg·kg^-1) in 12-week-old NIH male mice to prepare azoospermic model. Jinkui Shenqi Wan were administrated to azoospermia mice by gastric irrigation for 78 days (two spermatogenesis cycles) after 42 days of modeling. The testis total mRNAs of Jinkui Shenqi group and normal group were used to study gene expression profile by microarray. Result: The important fertility-related indicators such as body weight, reproductive organs and tissue mass index, sperm quality (motility, activity, concentration) and so on were improved significantly in azoospermia mice treated with Jinkui Shenqi Wan for 78 days. Jinkui Shenqi Wan played significant roles in recovering the spermatogenesis of testis in azoospermia mice. Compared with the normal group, microarray results showed that there were 804 genes (2.53%, 517 upregulation, 287 downregulation) with more than two times differential expression and 134 genes (0.42%, 127 upregulation, 7 downregulation) with four times differential expression, respectively. This suggested that the main role of Jinkui Shenqi Wan on recovery of spermatogenesis was positive and upregulation, negative and downregulation played secondery roles. Pathway analysis showed these genes expressed in: cell cycle, cytokine-cytokine receptor, Ecm receptors, Jak-Stat signaling pathway, Mapk signaling pathway, Notch signaling pathway, Toll-like receptor signaling pathway and Wnt signaling pathway. Conclusion: Testis spermatogesis of azoospermia mice can be recovered after administration of Jinkui Shenqi Wan by positive controlling genes related to cell proliferation through different pathways.