大鼠RBL-2H3细胞瞬时感受器电位M7通道参与细胞存活及凋亡

Role of transient receptor potential melastatin 7 in rat RBL-2H3 cells survival and apoptosis

背景:瞬时感受器电位M7是肥大细胞上重要的钙离子通道,但其在肥大细胞存活及凋亡过程中的作用仍未明确。目的:观察瞬时感受器电位M7通道对大鼠RBL-2H3细胞存活及凋亡的影响。方法:取对数生长期RBL-2H3细胞,①分别以50,100,200μmol/L瞬时感受器电位通道阻断剂2-APB进行干预,并以0.1%DMSO干预的细胞及正常培养的细胞作对照。②以瞬时感受器电位M7-siRNA反转录病毒载体转染RBL-2H3细胞,并设立空载体组和正常对照组。结果与结论:经过100,200μmol/L的2-APB作用72h后,RBL-2H3细胞数减少,吸光度值降低(P<0.05),细胞凋亡增多,早期凋亡率和总凋亡率增加(P<0.05);RBL-2H3细胞转染si-瞬时感受器电位M772h后,吸光度值降低(P<0.05),细胞凋亡增多,早期凋亡率和总凋亡率增加(P<0.05)。说明瞬时感受器电位M7通道参与了RBL-2H3细胞存活和凋亡过程。

BACKGROUND： Transient receptor potential melastatin 7 is an important calcium channel in mast cells. However, the role of transient receptor potential melastatin 7 in mast cell survival and apoptosis is unclear. OBJECTIVE： To investigate the role of transient receptor potential melastatin 7 in rat RBL-2H3 cells survival and apoptosis. METHODS： RBL-2H3 cells in logarithmic growth phase were intervened with 50, 100 and 200 μmol/L transient receptor potential channel blocker 2-APB. Cells intervened with 0.1% DMSO and normally cultured cells were used as control. RBL-2H3 cells were transfected with retrovirus vector of transient receptor potential melastatin 7-siRNA. Empty vector group and control group were constructed. RESULTS AND CONCLUSION： At 72 hour after treated with 100 and 200 μmol/L 2-APB, RBL-2H3 cell number was decreased. The optical density was decreased （P 0.05）. Cell apoptosis increased. Early stage apoptotic rate and total apoptotic rate increased （P 0.05）. At 72 hour after RBL-2H3 cells were transfected with transient receptor potential melastatin 7-siRNA, the optical density was decreased （P 0.05）. Cell apoptosis increased. Early stage apoptotic rate and total apoptotic rate increased （P 0.05）. These findings indicate that transient receptor potential melastatin 7 channel involve in the process of survival and apoptosis in RBL-2H3 cells.