何首乌饮延缓大鼠睾丸间质细胞衰老与调控胰岛素/胰岛素样生长因子-1信号通路的关系#br#

目的 探讨何首乌饮延缓大鼠睾丸间质（Leydig）细胞衰老与调控胰岛素/胰岛素样生长因子-1（IGF-1）信号通路的关系。方法 采用氧化损伤的方法建立大鼠Leydig细胞衰老模型。用流式细胞术检测各组细胞周期的变化。应用Real-time PCR、免疫荧光和Western blotting技术检测Leydig细胞胰岛素/IGF-1信号传导通路关键基因的表达水平,并运用胰岛素受体/IGF受体特异性抑制剂BMS-754807处理细胞,用流式细胞术检测细胞凋亡水平。结果 与正常组相比,衰老组β-半乳糖苷酶阳性细胞率可达60%（P＜0.05）,G1 期所占比重明显增加, S期所占比重明显减少（P＜0.05）。胰岛素/IGF-1信号传导通路关键基因表达变化：衰老组胰岛素受体（INSR）,胰岛素受体底物（IRS1）、IRS2、IGF1表达水平明显低于正常组（P＜0.05）,胰岛素样生长因子结合蛋白3（IGFBP3）的 mRNA表达明显高于正常组（P＜0.05）,何首乌饮可逆转上述现象。用 BMS-754807处理衰老的Leydig细胞后,Bcl-2 mRNA表达水平低于衰老组（P＜0.05）,用何首乌饮和BMS-754807同时处理衰老的Leydig细胞,细胞凋亡率高于何首乌饮组（P＜0.05）。结论 何首乌饮通过调控胰岛素/IGF-1信号传导通路延缓Leydig细胞衰老。

Relationship between the senescence of Leydig cells and the regulation of insulin/insuli-like growth factor-1 signaling pathway in rats delayed by Heshouwuyin#br#

Objective To investigate the relationship between the senescence of testicular Leydig cells and the regulation of insulin/insulin-like growth factor 1(IGF-1) signaling pathway in rats delayed by Heshouwuyin.Methods The oxidative damage technique was used to establish Leydig cell aging model. Flow cytometry was used to detect the changes of cell cycle in each group. Detection of mRNA and protein expression levels of key genes of insulin/IGF-1 signaling pathway in Leydig cells by Real-time PCR, immunofluorescence and Western blotting. Cells were treated with insulin receptor/IGF receptor specific inhibitor BMS-754807, flow cytometry was used to detect the apoptotic level of Leydig cells.Results Compared with the normal group, the proportion of β-galactosidase positive cells in the aging group was more than 60% (P<0.05), and the proportion of G1 phase increased significantly, while that of S phase decreased significantly (P<0.05); Changes in the expression of key genes of the insulin/IGF-1 signaling pathway: the expression levels of insulin receptor(INSR), insulin receptor substrate(IRS1), (IRS2), IGF-1 mRNA and protein in the aging group were significantly lower than those in the normal group（P＜0.05）, and the mRNA expression ofinsulin-like growth factor binding protein 3(IGFBP3) was significantly higher than in the normal group（P＜0.05）, Heshouwuyin could reverse the above phenomenon. After the treatment of senescent testicular Leydig cells with BMS-754807, the mRNA expression level of Bcl-2 was lower than that of the senescent group (P<0.05), and the cell apoptosis rate was higher than that of the senescent testicular Leydig cells treated with Heshouwuyin and BMS-754807at the same time (P<0.05).Conclusion Heshouwuyin delayed the senescence of testicular Leydig cells by regulating the insulin/IGF-1 signaling pathway.