靶向抑制NF-κB信号通过下调p16逆转睾丸间质干细胞衰老的研究

睾丸衰老所致的迟发性性腺功能减退症(LOH)严重影响老年男性的生活质量并与重大疾病相关，缺少理想疗法；我们前期证实睾丸间质干细胞(SLC)移植治疗LOH安全有效，但SLC来源限制了其临床应用。靶向逆转内源性干细胞衰老是抗衰老治疗的理想策略。预实验发现NF-κB信号与SLC衰老关系密切；体外激活NF-κB信号上调衰老特异基因p16表达；敲低IKKB可抑制NF-κB信号下调p16表达恢复老年小鼠SLC受损的自我更新和定向分化能力。据此，我们推测靶向抑制NF-κB信号可下调p16表达逆转SLC衰老。本研究拟通过RNA干扰和CHIP-seq等技术证实NF-κB信号在SLC衰老中的关键作用；明确NF-κB3与p16的结合位点；获取体内靶向NF-κB信号逆转内源性SLC衰老的可靠证据。研究结果将揭示SLC衰老的分子机制，促进SLC治疗的临床转化，并为LOH等睾丸衰老性疾病的治疗提供新思路。

Late-onset hypogonadism (LOH) affects the quality of life elderly males and contributes to other age-related diseases, such as diabetes and atherosclerosis. Testosterone replacement therapy is used to treat LOH, but with serious side effects. We had reported that stem Leydig cell (SLC) transplantation could rescue the testosterone production in rodent models of LOH. However, the shortage of SLCs limited their clinical application. Could rejuvenation aged SLCs broaden the resource of SLCs? We has demonstrated that TNF-α is up-regulated in the testis of 24-month old male mice. Moreover, hyper-activation NF-κB by TNF-α up-regulates the expression of p16 in young SLCs, which are derived from the testis of 3-month old male mice. And knockdown of IKKB by siRNA down-regulated the expression of p16 in aging SLCs, which are derived from the testis of 24-month old male mice. Accordingly, we hypothesized that inhibition NF-κB could rejuvenate SLCs by down-regulation the expression of p16. This study is aim to exploy techniques, such as CHIP-sequence and RNA interfering, to confirm the role of NF-κB signal in the senescence of SLC, to identify the binding sites between NF-κB3 and p16, to explore whether targeting NF-κB signal could improve therapeutic effects of transplanted aging SLC on hypogonadism, to further explore whether targeting NF-κB signal can rejuvenate endogenous senescent SLC to rescue dysfunction of aging testis. These findings will not only contribute to discover molecular mechanisms of the senescence of SLCs, to promote SLC clinical transplantation, but also shed new light on anti-aging of testis.