The Celluar Damage And Mechanism Of Hypoxia On Primary Sertoli Cells Of Rats

Objective:To construct testicular Sertoli cell hypoxic model in vitro. We plan to investigate the function and apoptosis of Sertoli cells in the male reproductive system in hypoxia state and speculate about the possible effect of hypoxia on the reproductive system. And we want to serch protective agent of hypoxia.Methods:On the basis of hypoxia of Sertoli cells induced by CoC12in vitro, we detect cell activity and spoptosis of Sertoli cells under hypoxia by MTT assay and flow cytometry instrument. And under different concentrations of CoC12simulation Sertoli cells activity changes. We detect apoptosis of Sertoli cells by real-time PCR, and chang of apoptotic effect molecules caspase-3by Western blot. And we detect secrete function of Sertoli cells under hypoxia.Results:We construct cell hypoxia model in vitro. Morphological changes, cell shrinkage happened under hypoxia. Activity of Sertoli cells under hypoxia reduced by dose-dependent, but not time-dependent. For Sertoli cells apoptosis is concentration dependent, and have time denpendece. Hypoxia induced by CoC12can result Sertoli cells apoptosis by relying on mitochondrial apoptosis pathway. And it was able to activate and promote MAPK signal pathway. Hypxoa can induce apoptosis of primary Sertoli cells by oxidative stress form. Hypoxia also can induce a function change of Sertoli cells. After that it can affect sperm production.Conclusion:We can construct cell hypoxia model of Sertoli cells in vitro successfully and we found that Sertoli cells have dose dependence of cell activity and apoptosis rate change. And MAPK signal pathway involved in apoptosis signal transduction. Apoptosis of activated by oxidative stress intrinsic apoptotic pathways.