This work aims to investigate the relationship between the senescence of testicular Leydig cells and the regulation of insulin/IGF-1 signaling pathway in rats delayed by Heshouwuyin.The oxidative damage technique was ...This work aims to investigate the relationship between the senescence of testicular Leydig cells and the regulation of insulin/IGF-1 signaling pathway in rats delayed by Heshouwuyin.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.The results showed that compared with the normal group,the proportion ofβgalactosidase 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 INSR,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 of IGFBP3 was significantly higher than in the normal group(P<0.05),Heshouwuyin can reverse the above phenomenon.展开更多
文摘This work aims to investigate the relationship between the senescence of testicular Leydig cells and the regulation of insulin/IGF-1 signaling pathway in rats delayed by Heshouwuyin.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.The results showed that compared with the normal group,the proportion ofβgalactosidase 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 INSR,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 of IGFBP3 was significantly higher than in the normal group(P<0.05),Heshouwuyin can reverse the above phenomenon.
