线粒体信号通路在热应激猪睾丸生精细胞凋亡中的作用机制研究

项目名称： 线粒体信号通路在热应激猪睾丸生精细胞凋亡中的作用机制研究

项目编号： No.31470124

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 贺俊平

作者单位： 山西农业大学

项目金额： 30万元

中文摘要： 哺乳动物精子发生受到包括激素和生长因子等多种因素的调控，热应激严重损害猪睾丸精子发生和精液品质，但其分子机制尚不清楚。对实验动物大鼠等的研究发现，热应激会导致生精细胞凋亡，线粒体信号通路是细胞凋亡的重要调节途径。热应激是否经由线粒体信号通路起始生精细胞凋亡，进而影响猪精子发生，未见研究报道。本研究以热应激猪睾丸组织为材料，用TUNEL法原位检测生精细胞凋亡的类型和数量；用透射电镜技术观察凋亡细胞凋亡小体和线粒体结构的变化；用QRT-PCR技术定量线粒体信号通路成员bcl-2, bax, cyt-c, caspase-3 mRNA的表达水平；用Western Blot技术分析线粒体信号通路凋亡相关蛋白的表达；用免疫组织化学技术确定线粒体信号通路凋亡相关蛋白在猪睾丸的组织细胞定位。以期探明线粒体信号通路在热应激致猪睾丸生精细胞凋亡中的作用机制，为热应激损伤猪精子发生提供分子靶标和理论依据。

中文关键词： 热应激；猪；睾丸；细胞凋亡；线粒体信号通路

英文摘要： Mammalian spermatogenesis was regulated by a variety of factors, including hormones and growth factors. The heat stress had detrimental effect on semen quality and spermatogenesis in the boar, but the mechanism remains unclear. Study of experimental animal rat found that heat stress could lead to the apoptosis of spermatogenic cells. Mitochondria signal pathway was one important regultor of apoptosis. Whether or not heat stress starting spermatogenic cell apoptosis by the mitochondria signal pathway, thereby affecting pig spermatogenesis, remains unclear as well. In this study, heat stress porcine testis was used as material. The apoptotic cell type and quantity was detected by TUNEL method. Apoptotic body and mitochondrial was observed by transmission electron microscopy. The expression of mitochondrial signal pathway members bcl-2, bax, cyt-c, caspase-3 were detected by QRT-PCR technique. The expression of mitochondrial pathway related proteins was analysed by Western Blot techinque. Localization of apoptotic related protein of mitochondrial signal pathway members in boar testis were conducted as well. The present study was designed to prove up the function mechanism of mitochondrial pathway in heat stress induced germ cell apoptosis in the boar, and to provide theoretical basis and molecular target for the ma

英文关键词： heat stress；boar；testis；apoptosis；mitochondria signal pathway