基于表观遗传学探讨益肾通络方对苯并(a)芘致生精细胞DNA损伤的防护机制

In recent years, the incidence of male infertility, recurrent abortion is rising, despite the complex reasons, unknown mechanism, it is closely related with the sperm DNA damaged or the stability of the genetic material affected. Environmental pollutants can invade the body, damage sperm DNA through the various ways such as breath, water,food, cause infertility, abortion, even by changing the sperm DNA epigenetic influence offspring reproductive system development, etc. Based on previous studies, research group put forward a hypothesis “Yishen Tongluo Decoction may have the effect of protect the spermatogenic cell DNA damage". To verify the hypothesis, we will establish animal and spermatagonial stem cells model infected by environmental pollutants benzo (a) pyrene(BaP), and treat with Yishen Tongluo Decoction. sperm (sperm of offspring)/spermatagonial stem cells will be conduct: Application of relevant experimental technique to detect DNA methylation level, DNA integrity, protein and mRNA expression of DNA repair protein (XRCC1, OGG1) and methylation transferase (DNMTs). It will provide theory support for traditional Chinese medicine protecting the genetic material damage caused by environmental pollution, as well as provides a new thought and method for TCM prevention and treatment of male infertility, male factors of recurrent miscarriage.

近年来，男性不育、复发性流产的发病率不断升高，尽管原因复杂，机制不明，但与精子DNA损伤，或精子DNA表观遗传学改变有着密切关系。环境污染物可通过呼吸、饮、食等途径侵袭机体，破坏精子DNA，或改变精子DNA表观遗传从而导致不育、流产或影响子代生殖系统发育等。课题组基于前期研究基础，提出“益肾通络方可能具有防护生精细胞DNA损伤的作用”的假说。通过建立环境污染物苯并(a)芘(BaP)染毒动物模型及精原干细胞模型，并用益肾通络方进行干预，对精子(及子代精子)/精原干细胞：应用相关实验技术检测DNA甲基化水平、DNA完整性、DNA修复蛋白(XRCC1、OGG1)、甲基化转移酶(DNMTs)蛋白及mRNA的表达等，验证假说。为中医药防护因环境污染物所致遗传物质的损伤，提供理论支持，同时也为中医药防治男性不育、复发性流产的男方因素的研究，提供新的思路和方法。

按照原计划完成研究任务。通过建立环境污染物苯并(a)芘(BaP)染毒动物模型及精原干细胞模型，并用益肾通络方进行干预，应用相关实验技术检测DNA甲基化水平、DNA完整性、DNA修复蛋白(XRCC1、OGG1)、甲基化转移酶(DNMTs)蛋白及mRNA的表达等。发现益肾通络方可对精子DNA直接损伤和表观遗传损伤两个层面对BaP染毒大鼠精子进行防护。在防护精子DNA直接损伤方面，发现其可能通过XRCC1或OGG1等DNA修复蛋白通路完成修复。在防护表观遗传损伤方面，发现其可能通过LncRNA通路防护由BaP造成等甲基化变化，而非通过DNMTs。共发表相关SCI文章2篇，中文文章6篇，申请国内专利4项。为中医药防护因环境污染物所致遗传物质的损伤，提供理论支持，同时也为中医药防治男性不育、复发性流产的男方因素的研究，提供新的思路和方法。