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目的探讨人参皂苷Rg1对1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinum,MPP+)诱导的PC12细胞凋亡是否具有保护作用。方法采用MPP+诱导的具有多巴胺能神经元特性的PC12细胞凋亡作为帕金森病(Parkinson disease,PD)的体外模型。实验分正常对照组、MPP+损伤组、人参皂苷Rg1(10、20、50μmol/L)3个浓度预处理组。用MTT法测定细胞活性,流式细胞术检测细胞凋亡,TUNEL法检测细胞凋亡断裂的DNA片段,蛋白质印迹法分析细胞色素C(cytochrome C,Cyt C)蛋白含量。结果 10、20、50μmol/L人参皂苷Rg1对MPP+诱导的PC12细胞具有一定的保护作用。与MPP+损伤组[(52±4.7)%]相比,10、20、50μmol/L人参皂苷Rg1预处理细胞活力分别上升为(64±3.4)%、(72±5.2)%、(83±6.2)%(P<0.05或P<0.01)。经流式细胞术检测,正常组、MPP+损伤组、人参皂苷Rg1预处理组(10、20、50μmol/L)细胞凋亡率分别为1.8%、44.5%、32.9%、21.1%和14.2%。人参皂苷Rg1预处理后,细胞断裂的DNA片段明显减少。另外,蛋白质印迹分析也显示人参皂苷Rg1可抑制MPP+诱导的Cyt C的过表达。结论人参皂苷Rg1对MPP+诱导的细胞凋亡具有浓度依赖性的保护作用,其保护机制可能与下调线粒体内Cyt C的过表达有关。
Objective To investigate whether ginsenoside Rg1 can protect PC12 cells from apoptosis induced by 1-methyl-4-phenylpyridinum (MPP +). Methods Apoptosis of PC12 cells with dopaminergic neuron induced by MPP + was used as an in vitro model of Parkinson disease (PD). The experiment was divided into 3 groups: normal control group, MPP + injury group and ginsenoside Rg1 (10, 20, 50μmol / L). Cell viability was measured by MTT assay, apoptosis was detected by flow cytometry, DNA fragmentation was detected by TUNEL method, and protein content of cytochrome C (Cyt C) was analyzed by Western blotting. Results 10, 20 and 50μmol / L ginsenoside Rg1 could protect PC12 cells induced by MPP +. Compared with MPP + injury group [(52 ± 4.7)%], cell viability of 10, 20 and 50μmol / L ginsenoside Rg1 pretreatment increased to (64 ± 3.4)%, (72 ± 5.2)% and )% (P <0.05 or P <0.01). The apoptosis rate of normal group, MPP + injury group and ginsenoside Rg1 pretreatment group (10, 20 and 50μmol / L) were 1.8%, 44.5%, 32.9%, 21.1% and 14.2% respectively by flow cytometry. After pretreatment with ginsenoside Rg1, the DNA fragmentation of the cells was significantly reduced. In addition, Western blot analysis also showed that ginsenoside Rg1 inhibits MPP + -induced Cyt C overexpression. Conclusion Ginsenoside Rg1 has a concentration-dependent protective effect on MPP + -induced apoptosis, and its protective mechanism may be related to down-regulation of Cyt C overexpression in mitochondria.