基于C-KIT/PI3K/AKT/P70S6K信号通路研究调肝中药治疗VC性不育的作用机理

精索静脉曲张（VC）是引起男性不育的第一大原因，前国自然项目研究发现，VC大鼠睾丸干细胞因子（SCF）及其受体C-KIT表达发生显著改变，在调肝中药干预后，SCF/C-KIT表达恢复，生精功能改善，表明调肝中药可能是通过干预SCF/C-KIT表达而改善生精功能的。进一步我们将探索C-KIT下游通路的介导作用。本研究拟转染C-KIT低表达病毒降低生精细胞C-KIT表达，然后与支持细胞共培养，通过添加PI3K/AKT激活剂及调肝中药含药学清，分析PI3K/AKT/P70S6K通路介导调肝中药治疗VC的作用；在VC大鼠睾丸内感染C-KIT过表达病毒载体、注射PI3K/AKT激活剂及调肝中药灌胃，测定SCF/C-KIT及PI3K/AKT/P70S6K分子表达，阐明VC不育是通过SCF/C-KIT介导PI3K/AKT/P70S6K通路，导致生精障碍，而调肝中药是通过调节此通路改善VC生精过程。

Varicocele (VC) is the first reason for causing male infertility. Our previous program found that the expression of stem cell factor (SCF) and C-KIT receptor changed obviously in rat with VC, and spermatogenesis improved and the expression of SCF and C-KIT recovered after intragastric administration of Liver-regulating traditional medicine(LRTM), which suggested that LRTM can improve spermatogenesis by intervene the expression of SCF/C-KIT. Next, the role of mediation of C-KIT for its downstream signal path will be focused on. In this research, transfection of virus vector to decrease the expression of C-KIT in germ cells, and co-culture with Sertoli cells. The mediation effect of PI3K/AKT/P70S6K pathway will be analyzed by adding the angonist of PI3K/AKT and serum including LRTM to cells. In vivo, overexpression of C-KIT, injection of the angonist of PI3K/AKT and intragastric administration of LRTM in the testis of VC rats, the expression of SCF/C-KIT and PI3K/AKT/P70S6K will be detected to explore SCF/C-KIT mediate the pathway of PI3K/AKT/P70S6K to result spermatogenic failure in VC causing male infertility, and LRTM improve the spermatogenesis by regulating the pathway.