Annexin 5刺激大鼠睾丸Leydig细胞后的差异表达蛋白的研究

Differentially expressed proteins in the process of annexin 5 stimulating testosterone secretion from rat Leydig cells

目的:研究annexin 5作用于大鼠睾丸Leydig细胞后的差异表达蛋白。方法:原代培养大鼠睾丸Leydig细胞,1 nmol/L的annexin 5处理Leydig细胞24 h,提取细胞蛋白用二维电泳分离蛋白并进行比较。选择与对照组有明显差异表达的蛋白点,进行质谱分析。结果:获得了分辨率和重复性均很好的差异蛋白图谱。筛选出的显著差异表达的50个蛋白点,共有36个蛋白点被成功鉴定,其中annexin 5处理组中高表达的为23个,低表达的为13个。结论:初步建立了annexin 5作用于大鼠睾丸Leydig细胞后的差异蛋白谱,筛选并鉴定出的这些蛋白质可能是影响睾丸Leydig细胞合成睾酮过程中的关键蛋白,研究结果为阐明annexin 5调控大鼠睾丸Leydig细胞分泌睾酮的具体机制奠定了基础。

Objective： To study the differentially expressed proteins in the process of annexin 5 stimulating testosterone secretion in cultured rat Leydig cells.Methods： Primary rat Leydig cells were cultured in vitro and treated with annexin 5 at the concentration of 1 nmol/L for 24 hours,and the cell proteins were extracted to be compared by two-dimensional gel electrophoresis（2-DE）.The differentially expressed protein spots were selected to be analyzed by mass spectrometry.Results： We obtained electrophoresis profiles with high resolution and reproducibility,found 50 differentially expressed protein spots,and identified 36 by mass spectrometry,of which 23 were overexpressed and 13 underexpressed in the Leydig cells treated with annexin 5.Conclusion： Differentially expressed protein profiles were established in the process of annexin 5 stimulating testosterone secretion in cultured rat Leydig cells,and identified the key role of these proteins in testosterone secretion.Our findings might be helpful to illuminate the mechanism of annexin 5 regulating testosterone secretion in rat Leydig cells.