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

LI Shao-Hua; JIANG Li-Ping; LIU Liang-Yun; LIU Wen-Qing; ZHU Juan; NIU Si-Yun; QI Feng

doi:10.16098/j.issn.052-1356.2020.05.019

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (5) : 751-757. DOI: 10.16098/j.issn.052-1356.2020.05.019

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

LI Shao-hua² JIANG Li-ping¹ LIU Jing-yun¹ LIU Wen-qing¹ ZHU Juan¹ NIU Si-yun^1* QI Feng3*

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Abstract

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 of insulin-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.

Key words

Heshouwuyin / Testicular Leydig cells / Insulin/insulin-like growth factor-1 signaling pathway / Western blotting / Rat

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LI Shao-hua JIANG Li-ping LIU Jing-yun LIU Wen-qing ZHU Juan NIU Si-yun QI Feng. Relationship between the senescence of Leydig cells and the regulation of insulin/insuli-like growth factor-1 signaling pathway in rats delayed by Heshouwuyin[J]. Acta Anatomica Sinica. 2020, 51(5): 751-757 https://doi.org/10.16098/j.issn.052-1356.2020.05.019

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