海龙黄精散通过抑制微波辐射诱导的睾丸支持细胞过度凋亡调控精原干细胞增殖与分化的机制研究

睾丸是微波的靶器官，微波辐射可致睾丸生精障碍。微波可直接损伤生精细胞和支持细胞，而受损支持细胞在生精障碍中的作用机制尚不清楚。我们前期研究发现，海龙黄精散可治愈微波辐射性睾丸生精障碍，同时受损的支持细胞也得以修复，但未对受损支持细胞修复与生精障碍改善的相关性进行探讨。今推断海龙黄精散可通过抑制微波诱导的支持细胞过度凋亡调控SSCs增殖与分化。本项目以雄小鼠为研究对象，建立微波辐射不育模型，用海龙黄精散治疗。电镜观察支持细胞及生精细胞变化；IF法检测Bax、Bcl-2、caspase-3表达；ELISA法检测脂质过氧化物；FQ-PCR法检测睾丸GDNF、SCF、BMP4的mRNA表达水平，蛋白免疫印迹法和IF法检测PI3K和p-Akt的表达与定位。研究微波损伤的支持细胞对SSCs增殖与分化影响机制及海龙黄精散治疗机理，为微波辐射男性不育提供新疗法，为补肾理论提供现代生物学诠释。

The testicle is the target organ of microwave. Microwave can directly damage spermatogenic cells and sertoli cells, but the mechanism of damaged sertoli cells in spermatogenesis dysfunction is not clear. In our previous study, we found that Hai Long Huang Jing Powder could cure the testicular spermatogenesis dysfunction caused by microwave radiation and repair the damaged sertoli cells, but we did not discuss the correlation between the repair of the damaged sertoli cells and the improvement of the spermatogenesis dysfunction. We can infer that Hai Long Huang Jing Powder can regulate the proliferation and differentiation of SSCs by inhibiting the over-apoptosis induced by microwave. In this study, male mice will be used to establish a microwave radiation infertility model and be treated with Hai Long Huang Jing Powder. The changes of sertoli cells and spermatogenic cells will be observed by electron microscopy. The expressions of Bax, bcl-2 and caspase-3 will be detected by IF method. Lipid peroxide will be detected by ELISA method. The mRNA expression levels of GDNF, SCF and BMP4 in testis will be detected by FQ-PCR method, and the expression and localization of PI3K and p-Akt will be detected by western blot and IF method. The study on the effect mechanism of the sertoli cells damaged by microwave on the SSCs proliferation and differentiation and the treatment mechanism of Hai Long Huang Jing Powder will provide a new treatment for male infertility caused by microwave radiation and a modern biological interpretation for the theory of supplementing the kidney.