补肾中药通过AMPK-mTOR通路提高肾精亏虚大鼠睾丸间质细胞自噬能力

Kidney-tonifying Chinese medicine improves autophagy of Leydig cells in rats with kidney essence deficiency through AMPK-mTOR pathway

为了探究补肾中药提高肾精亏虚大鼠睾丸间质细胞自噬能力的机制,采用单因素电刺激法建立肾精亏虚大鼠模型,免疫组化及Western blot检测各组睾丸细胞自噬标志蛋白Beclin1、p62、LC3B表达水平以及AMPK和mTOR表达变化.基于肾精亏虚与衰老的密切联系,通过自由基氧化损伤法建立睾丸间质细胞(Leydig细胞)衰老模型,联合AMPK抑制剂,探究何首乌饮激活细胞自噬的分子机制.结果显示:何首乌饮可上调睾丸组织Beclin1蛋白、LC3B表达,下调p62蛋白表达,促进AMPK磷酸化并抑制mTOR激活(P<0.05);细胞实验表明,何首乌饮可通过激活AMPK-mTOR信号通路提高Leydig细胞自噬能力(P<0.05),促进肾精亏虚大鼠睾丸Leydig细胞自噬进程.

In order to explore the mechanism of kidney-tonifying Chinese medicine in improving autophagy of testicular mesenchymal cells in rats with kidney-essence deficiency,a rat model of kidney-essence deficiency was established by single-factor electrical stimulation.Immunohistochemistry and Western blot were used to detect the expression levels of autophagy marker proteins Beclin1,p62,LC3B,AMPK and mTOR in testicular cells of each group.Based on the close relationship between kidney essence deficiency and aging,Leydig cell aging model of testicular mesenchymal cells was established by free radical oxidative damage method,and AMPK inhibitor was combined to explore the molecular mechanism of Heshouwuyin activating autophagy.The results showed that Heshouwuyin could up-regulate the expression of Beclin1 protein and LC3B in testicular tissue,down-regulate the expression of p62 protein,promote AMPK phosphorylation and inhibit mTOR activation(P<0.05).Cell experiments showed that Heshouwuyin could affect the autophagy ability of Leydig cells by activating AMPK-mTOR signaling pathway(P<0.05).Therefore,Heshouwuyin may promote the autophagy process of Leydig cells in rats with kidney essence deficiency by activating AMPK-mTOR signaling pathway.