原儿茶醛对环磷酰胺致小鼠精子发生异常的保护作用及机制研究

DJ-1 protein is closely related with male infertility, and protocatechualdehyde interacts with DJ-1 protein. Our preliminary study showed that protocatechualdehyde inhibited cyclophosphamide-induced mouse spermatogenesis abnormity, with the mechanisms to be studied. This project is aimed to explore the effects and mechanisms of protocatechualdehyde on cyclophosphamide-induced mouse spermatogenesis abnormity, with the research core of spermatogenesis and the main line of blood-testis barrier→spermatogenesis→sperm. The effects of protocatechualdehyde on blood-testis barrier (the structure of blood-testis barrier, tightly connected protein expression, cytoskeleton protein expression and MAPK-ERK signal pathway), spermatogenesis (the structure of spermatogenic tubules, proportion of different types of spermatocytes, spermatogonia marker c-kit, apoptosis of spermatocytes and DJ-1-PI3K-Akt-mTOR signal pathway) and sperm (sperm density and motility, sperm DNA damage, sperm ATP content and sperm DJ-1 protein expression) will be studied. This project will help obtaining potential drug for male infertility and potential drug antagonizing male reproductive toxicity of cyclophosphamide, with significant scientific research meannings and extensive social application prospects.

DJ-1蛋白与男/雄性不育密切相关，而原儿茶醛能与DJ-1蛋白发生相互作用。我们的前期研究结果提示原儿茶醛对环磷酰胺致小鼠精子发生异常具有保护作用，但其机制有待于研究。本项目拟进一步深入系统研究原儿茶醛对环磷酰胺致小鼠精子发生异常的保护作用及其机制：以精子发生为核心，以血睾屏障→精子发生→精子为主线，研究原儿茶醛对血睾屏障（血睾屏障结构、紧密连接蛋白表达、细胞骨架蛋白表达以及MAPK-ERK信号通路）、精子发生（生精小管结构、不同类型生精细胞比例、精原细胞标志物c-kit表达、生精细胞凋亡以及DJ-1-PI3K-Akt-mTOR信号通路）和精子（精子的密度和活力、精子DNA损伤、精子ATP含量以及精子DJ-1蛋白表达）的影响。本项目的顺利实施有助于研发新的治疗男性不育症的潜在药物，并获得拮抗环磷酰胺的男性生殖毒性的潜在药物，具有重要的科学研究意义和广泛的社会应用前景。

近年来，由于生活方式的改变和环境因素的影响，不育夫妇越来越多，其中50%是由男性因素导致的。DJ-1蛋白与男/雄性不育密切相关，而原儿茶醛能与DJ-1蛋白发生相互作用。我们推测并发现了原儿茶醛对雄性不育动物具有治疗作用，但其机制有待于研究。本项目以精子发生为核心，以血睾屏障→精子发生→精子为主线，进一步深入系统地研究了原儿茶醛对环磷酰胺致小鼠精子发生异常的保护作用及其机制。研究结果显示，原儿茶醛能上调紧密连接蛋白Occludin、ZO-1和细胞骨架蛋白F-actin的表达，故原儿茶醛对环磷酰胺导致的血睾屏障损伤有保护作用。原儿茶醛还能保护睾丸形态、使生精小管结构维持正常，上调精原细胞标志物c-kit的表达，并减少生精细胞凋亡，提示原儿茶醛对精子发生具有保护作用，进一步研究发现其机制与激活睾丸DJ-1-PI3K-Akt-mTOR信号通路相关。原儿茶醛还能减少精子DNA损伤并上调精子DJ-1蛋白表达。因此，我们认为原儿茶醛通过对血睾屏障的保护作用，促进精子发生，并保护精子损伤，是潜在的治疗男性不育症的药物，并能预防和治疗化疗药环磷酰胺的生殖毒性，具有一定的应用前景。