从AMPK/SIRT1调节自噬途径研究衰老睾丸Sertoli细胞功能衰退机制与淫羊藿总黄酮的干预

我们前期结题基金研究显示，增龄过程中睾丸生精障碍与Sertoli细胞功能衰退直接相关，且淫羊藿总黄酮可显著改善衰老睾丸生精功能，但Sertoli细胞功能衰退发生机理与淫羊藿总黄酮干预机制仍有待研究。细胞自噬功能下降是机体衰老的主要原因，新近我们发现，衰老大鼠睾丸组织中自噬调节分子AMPK、SIRT1表达下调，提示自噬能力下降可能是Sertoli细胞功能衰退的主要机制，而淫羊藿总黄酮可能通过AMPK/SIRT1通路改善Sertoli细胞自噬。本项目拟采用自然衰老大鼠与Sertoli细胞模型，运用免疫荧光、Western blot、siRNA等技术，首先研究AMPK/SIRT1通路介导自噬变化与Sertoli细胞功能衰退的关系，随后研究淫羊藿总黄酮对AMPK/SIRT1通路与自噬调节作用，进而探讨衰老睾丸Sertoli细胞功能衰退与淫羊藿总黄酮干预机制，为临床防治衰老睾丸生精障碍提供科学依据。

Our studies from completed National Natural Science Foundation of China have demonstrated that spermatogenic disorder with increasing age is directly related to the decline of Sertoli cell function, and total flavonoids of Epimedium (TFE) could significantly ameliorate the spermatogenic function of aging testis. However, the molecular mechanism of Sertoli cell dysfunction during aging and the intervention mechanism of TFE remain unclear. The decreased autophagy function is the main cause of aging body, and we found that autophagy-related molecules such as AMPK and SIRT1 expression levels were down-regulated in aging testis. These data suggest that the decreased autophagy ability may be the main mechanism leading to the decline of Sertoli cell function, while TFE improves Sertoli cell autophagy via AMPK/SIRT1 signal pathway. Thus, in natural aging rats and Sertoli cell model, we propose to clarify the relationship between autophagy mediated by AMPK/SIRT1 signal pathway and the decline of Sertoli cell function using immunofluorescence, Western blot and siRNA techniques. And we will investigate the regulatory effect of TFE on AMPK/SIRT1 signal pathway and autophagy, and then explore the mechanism of functional decline of Sertoli cells and intervention mechanism of TFE in aging testis. It will provide a necessary scientific basis for prophylaxis and therapy of spermatogenic disorder of aging testis.