迟发性性腺功能减退症（LOH）严重影响中老年男性健康，睾丸间质干细胞（SLCs）衰老，无法有效补充合成睾酮的睾丸间质细胞（LCs）是可能的发病机制。氧化应激损伤在干细胞衰老中扮演了重要角色，申请人前期研究证实中间丝蛋白Nestin是SLCs的特异标志，在SLCs中下调Nestin的表达导致抗氧化体系关键蛋白Nrf2表达下降，氧化应激损伤增强，SLCs衰老增加；进一步通过质谱分析发现Nestin能与Nrf2降解调节蛋白Keap1直接结合，提示Nestin可能通过Keap1-Nrf2抗氧化通路参与调控SLCs衰老，但其具体调控机理远未阐明。据此，本研究拟利用超分辨显微镜、条件性基因敲除小鼠等方法，深入解析Nestin-Keap1-Nrf2环路在SLCs稳定维持与衰老中的重要作用，揭示Nestin如何通过与Keap1竞争性结合调控Nrf2的降解与转录活性，为发现LOH治疗的新靶点提供实验依据。

Late-onset hypogonadism (LOH) seriously impairs the health of elderly men. With the senescence of stem Leydig cells (SLCs), the capability of SLCs to generate endogenous testosterone-producing Leydig cells (LCs) is gradually diminished, which is an important cause of LOH. Oxidative stress injury plays an important role in stem cell senescence. Previous studies of the applicant have confirmed that Nestin is a specific marker of SLCs. Down-regulation of Nestin can decrease the expression of Nrf2, a key protein in the antioxidant system, thus exposing SLCs to oxidative stress injury and accelerating the senescence progress. Furthermore, mass spectrometry data analysis shows that Nestin can bind to Keap1, the Nrf2 degradation regulatory protein, suggesting that Nestin may regulate SLCs senescence through the Keap1-Nrf2 antioxidant pathway, but Nestin’s specific regulatory mechanism remains unknown. Accordingly, this study is intended to analyze the significant role of Nestin-Keap1-Nrf2 loop in the stabilization and senescence of SLCs by means of super-resolution microscopy and conditional knockout mice, and uncovers the mechanism of that Nestin regulates the degradation and transcriptional activity of Nrf2, which will provide experimental basis for new targets of LOH therapy.