瓜子金皂苷己对MPP~+诱导PC12细胞凋亡的保护作用

目的观察瓜子金皂苷己(polygalasaponin F,PS-F)对1-甲基-4-苯基-吡啶离子(1-methyl-4-phenylpyridinium,MPP+)诱导的PC12细胞损伤的影响,并且探讨其作用机制。方法 MTT法检测细胞存活率,Annexin V/PI染色流式细胞术检测PC12细胞凋亡,JC-1染色倒置显微镜检测细胞线粒体膜电位(mitochondrial membrane potential,MMP),Western blot检测Caspase-3蛋白的水平。结果 500μmol.L-1MPP+作用PC12细胞48 h,能明显抑制细胞生长(P<0.01),诱导细胞发生凋亡,同时降低MMP,增加活性Caspase-3的蛋白水平。同时给予不同浓度PS-F处理,PC12细胞存活率增加(P<0.01);凋亡细胞量减少;MMP增高;活性Caspase-3蛋白水平降低(P<0.01)。结论 PS-F能抑制MPP+诱导的PC12细胞的凋亡,其作用机制可能与维持线粒体正常膜电位,稳定线粒体功能,降低活性Caspase-3蛋白水平有关。

The protective effect of polygalasaponin Fagainst MPP~+ induced PC12 cellular apoptosis

Aim To observe the effects and mechanism of polygalasaponin F(PS-F) on PC12 cellular apoptosis induced by 1-methyl-4-phenylpyridinium ion(MPP+).Methods MTT assay was used to measure the viability of the PC12 cells and flow cytometry was used to analyze the apoptotic rate of PC12 cells.The expression of active Caspase-3 in PC12 cells was detected by Western blot.In addition,the mitochondrial membrane potential was determined by JC-1 staining inverted microscope.Results When exposed to 500 μmol·L-1 MPP+ for 48 h,the viability and the mitochondrial membrane potential of PC12 cells decreased significantly,and the apoptosis rate and the expression of active Caspase-3 were increased significantly.Treatment with PS-F,the cell viability(P<0.01) and the MMP were significantly increased,the apoptosis rate and the expression of active Caspase-3(P<0.01) were significantly decreased compared with MPP+ group.Conclusion PS-F protects PC12 cells against apoptosis induced by MPP+ and the mechanism may be related to ameliorate the mitochondrial dysfunction and decrease the expression of active Caspase-3.