人参皂苷Rg1对1-甲基-4-苯基吡啶离子诱导的PC12细胞凋亡的保护作用
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Neuroprotective effects of ginsenoside Rg1 on 1-methyl-4-phenylpyridinum-induced apoptosis in PC12 cells
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    摘要:

    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、50 μmol/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、50 μmol/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、50 μmol/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、50 μmol/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。

    Abstract:

    ObjectiveTo explore the neuroprotective effect of ginsenoside Rg1 against PC12 cell apoptosis induced by 1-methyl-4-phenylpyridinum(MPP+). MethodsMPP+-induced apoptosis in PC12 cells, with the characteristics of dopaminergic neuron, were taken as the model of Parkinson disease in vitro. The cells were divided into control group, MPP+ group and 3 ginsenoside Rg1 pretreatment groups (concentrations 10, 20, and 50 μmol/L). MTT assay was used for detecting the cell viability, FCM for apoptosis ratio, TUNEL enzyme labelling for DNA fragment of the cell nuclear, and Western blotting analysis for cytochrome C protein.ResultsGinsenoside Rg1 (10, 20, and 50 μmol/L) showed protective effect against MPP+-induced PC12 cells injury. Compared with MPP+-treated cells(\[52±4.7\]%), pretreatment with 10, 20, and 50 μmol/L ginsenoside Rg1 increased the cell viability to (64±3.4)%, (72±5.2)% and (83±6.2)%, respectively (P<0.05 or P<0.01). FCM analysis indicated that apoptosis rates decreased by ginsenoside Rg1 pretreatment, with the apoptosis rates in the control, MPP+ and 3 ginsenoside Rg1 groups (10, 20, 50 μmol/L) being 1.8%, 44.5%, 32.9%, 21.1% and 14.2%, respectively. We also found that ginsenoside Rg1 pretreatment greatly decreased DNA fragment of PC12 cells. Western blotting analysis indicated that the cytochrome C was depressed by the ginsenoside Rg1 pretreatment. ConclusionGinsenoside Rg1 can protect PC12 cells against MPP+-induced apoptosis in a concentration-dependent manner, which may be closely related to down-regulation of cytochrome C over-expression in the mitochondria.

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  • 收稿日期:2011-04-28
  • 最后修改日期:2011-06-02
  • 录用日期:2011-08-29
  • 在线发布日期: 2011-09-22
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