苯并[a]芘（B[a]P）是一种广泛存在的环境内分泌干扰物，对雄性生殖系统具有危害作用。研究表明B[a]P可损伤睾丸间质细胞并影响睾酮合成过程造成雄性生殖损害，但其中所涉及的靶细胞器和具体的分子机制尚不清楚。我们前期研究发现B[a]P可诱导小鼠睾酮间质细胞株TM3线粒体和内质网损伤，且睾酮合成过程需要线粒体和内质网的参与，推测线粒体和内质网可能是B[a]P致睾酮合成障碍的重要损伤靶点。本项目拟建立B[a]P的活性代谢产物BPDE染毒原代小鼠睾酮间质细胞模型和B[a]P染毒动物模型，采用电镜、western blot、Q-PCR、流式细胞术、ELISA等方法检测睾酮水平、线粒体损伤、内质网应激和CHOP介导的PGC-1α通路关键蛋白的变化，旨在阐明线粒体损伤和内质网应激在B[a]P致睾丸间质细胞睾酮合成障碍中的作用及分子机制，为深入理解B[a]P致雄性生殖损伤的毒理机制提供新的思路。

Benzo[a]pyrene (B[a]P) is a widely existed environmental endocrine disruptor, which is harmful to male reproductive system. Previous studies have shown that B[a]P can damage Leydig cells and affect the process of testosterone synthesis, cause male reproductive damage, however, the target organelles and underlying molecular mechanisms involved are still unclear. Our previous study found that B[a]P can destroy the mitochondria and endoplasmic reticulum integrity of the mouse Leydig TM3 cell line, and the process of testosterone biosynthesis requires the participation of mitochondria and endoplasmic reticulum. From this, we speculated that mitochondria and endoplasmic reticulum may be important targets of testosterone synthesis disorder induced by B[a]P. In order to investigate the roles and molecular mechanisms of mitochondrial damage and endoplasmic reticulum stress in testosterone biosynthesis in Leyding cells exposed to B[a]P, the present study plans to establish in vitro model of BPDE-exposed the purified mouse Leydig cells and in vivo model of B[a]P-exposed mice, Testosterone concentration, mitochondrial ultrastructure, mitochondrial membrane potential, ATP level, mitochondrial DNA copy number, and expression of CHOP, PGC-1α signaling pathway related proteins, testosterone biosynthesis related proteins were measured through ELISA, transmission electronmicroscope, flow cytometry , ATP detection kit, Q-PCR and western blot, respectively. These findings will provide new ideas for further understanding the toxic mechanism of B[a]P-induced male reproductive damage.