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目的探讨线粒体相关基因Cox7a2编码蛋白对睾酮生成的影响。方法构建Cox7a2-EYFP-N1荧光表达载体,鉴定后转染TM3小鼠睾丸Leydig细胞,荧光显微镜观察转染效率和蛋白表达。ELISA的方法测定上清液中睾酮生成水平,CCK-8细胞增殖和生长测定试剂盒检测Cox7a2-YFP融合蛋白对细胞增殖状态的影响。结果 Cox7a2-YFP在TM3细胞中表达后抑制LH诱导的睾酮分泌,细胞增殖试验显示,Cox7a2融合蛋白表达后能抑制TM3细胞的增殖。结论线粒体定位蛋白Cox7a2能抑制TM3细胞睾酮合成,与Cox7a2对细胞增殖有抑制作用有关。
Objective To investigate the effect of mitochondria-related gene Cox7a2 on testosterone production. Methods Cox7a2-EYFP-N1 fluorescent expression vector was constructed and transfected into Leydig cells of TM3 mice after transfection. The transfection efficiency and protein expression were observed under fluorescence microscope. ELISA method was used to measure the level of testosterone in the supernatant. The effect of Cox7a2-YFP fusion protein on cell proliferation was tested by CCK-8 cell proliferation and growth assay kit. Results Cox7a2-YFP inhibited the LH-induced testosterone secretion in TM3 cells. Cell proliferation assay showed that Cox7a2 fusion protein inhibited the proliferation of TM3 cells. Conclusion Mitochondrial localization protein Cox7a2 can inhibit testosterone synthesis in TM3 cells, which is related to the inhibitory effect of Cox7a2 on cell proliferation.