RNA干扰PPT1基因表达对小鼠Sertoli细胞功能的影响

摘要

Objective To investigate the function of PPT1 gene in mose sertoli cells. Methods The expression of PPT1 gene in Sertoli cell line TM4 was inhibited using RNAi technique. Viability of transfected cells was measured by MTS. Meanwhile, cell apoptosis was detected by Flow Cytometry. The cells were stained by Mito Tracker Red and their mitochondrial activity was observed under the laser scanning confocal microscope.The ultrastructure of cell mitochondria was observed by transmission electron microscopy (TEM). Results The inh ibition of PPT1 gene expression decreased cell viability and mitochondrial activity, and resulted in the ultrastructural changes of the mitochondria as well as enhanced cell apoptosis rate. Conclusion PPT1 plays a critical role in maintaining the structure and function of Sertoli cells.%目的：探索PPT1基因在小鼠支持细胞（Sertoli细胞）中的功能。方法采用RNA干扰技术下调小鼠Sertoli细胞株TM4中PPT1基因的表达，并运用MTS法检测细胞活力，Annexin V染色结合流式细胞技术检测细胞凋亡，Mito Tracker Red染料染色细胞，并在激光共聚焦显微镜下观察线粒体且测得荧光强度以检测线粒体活性，透射电镜观察线粒体超微结构变化。结果抑制小鼠TM4细胞中PPT1基因的表达可降低其细胞活力和线粒体活性，导致线粒体超微结构发生改变，并增加细胞凋亡率。结论 PPT1基因对维持小鼠Sertoli细胞的功能具有重要作用。