睾丸是雄性激素产生的重要器官，性激素与DNA甲基化和衰老进程关系密切。本研究在祖国医学“肾主生殖”理论指导下，结合肾精盈亏与性腺和衰老的相关性，前期基因芯片筛选发现，补肾中药何首乌饮可使衰老大鼠睾丸组织表观遗传基因、细胞凋亡信号通路、胰岛素网络节点等相关的912个基因发生改变，且何首乌饮可促进睾丸间质细胞分泌睾酮。并且通过荧光定量PCR、荧光免疫和免疫印迹方法，证明了胰岛素网路节点关键基因IGF1、IR、IRS1、IRS2和SOCS3等受何首乌饮调控。因此，我们推测补肾中药何首乌饮通过调控表观遗传基因和胰岛素网络节点，抑制生精细胞凋亡，延缓肾精亏虚雄性大鼠生殖衰老。本项目以“房劳加惊恐”所致的肾精亏虚雄性大鼠为研究对象，运用DNA甲基化芯片及甲基化特异PCR技术结合荧光免疫染色等方法，旨在阐明补肾中药在肾精亏虚大鼠生殖衰老过程中的DNA甲基化机制，丰富“肾主生殖”中医藏象理论的科学内涵。

Testis is an important organ which secretes male hormones and it’s closely associated with geriatric diseases and infertility.Under the guidance of Chinese medicine theory ‘the kidney controlling reproduction’,combining with the correlation of kidney essence,aging and gonadal axis,we used gene chip method found that: Heshouwuyin could influence 912 genes expression in aging rats testes,these genes were related to epigenetics,cell apoptosis and insulin signaling pathway,in addition,Heshouwuyin could promote the Leydig cells ability in secreting testosterone,we also found that several key genes of insulin signaling pathway,including IGF1,IR, IRS1, IRS2 and SOCS3 were regulated by Heshouwuyin via fluorescent quantitative PCR,immunofluorescence and western blot method.Therefore,we hypothesized that Chinese kidney-tonifying drugs -Heshouwuyin could inhibit spermatogenic cells apoptosis and delay senescence by regulating the epigenetic gene and insulin network nodes.In this project,sexual strain and panic male rats work as kidney essence deficient model,we plans to explore the DNA methylation mechnism of Chinese kidney-tonifying drugs in delaying reproductive aging of kidney essence deficient rats by means of DNA methylation microarray,methylation specific PCR and immunofluorescence technology,so as to enrich TCM Zangxiang theory:the kidney controlling reproduction.