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目的初步研究不同浓度的五味子甲素(Schisandra A,Sch A)对1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium ion,MPP~+)引起的SH-SY5Y细胞凋亡的影响。方法体外培养人神经母细胞瘤SH-SY5Y细胞,1 mmol/L MPP~+染毒同时分别给予1、3和5μmol/L Sch A处理48 h,Muse~(TM)细胞分析仪检测凋亡细胞数的变化;hoechst33342染色后激光共聚焦显微镜进一步观察各组细胞形态的改变;Western-blot检测各组细胞中cleaved caspase-3蛋白表达水平的变化。结果与对照组相比,1 mmol/L MPP~+染毒48 h可引起SH-SY5Y细胞凋亡细胞数明显增多;1、3和5μmol/L Sch A干预可明显抑制1mmol/L MPP~+诱导的SH-SY5Y细胞凋亡,且随着Sch A浓度的升高,凋亡细胞数明显减少。Western-blot实验结果显示,MPP~+组细胞中cleaved caspase-3蛋白表达水平明显升高,而Sch A干预组cleaved caspase-3蛋白表达明显降低(P<0.05)。结论 1 mmol/L MPP~+可诱导SH-SY5Y细胞凋亡,而一定浓度的Sch A能有效抑制MPP~+引起的SH-SY5Y细胞凋亡,且该作用可能是通过抑制caspase-3蛋白活化实现的。
Objective To study the apoptosis of SH-SY5Y cells induced by 1-methyl-4-phenylpyridinium ion (MPP ~ +) with different concentrations of Schisandra A (SchA) Impact. Methods Human neuroblastoma SH-SY5Y cells were cultured in vitro. The cells were treated with 1 and 5 μmol / L SchA for 48 h with 1 mmol / L MPP ~ +. The apoptotic cells were detected by Muse ~ (TM) The changes of cell morphology were observed by laser scanning confocal microscopy after hoechst33342 staining. The expression of cleaved caspase-3 protein in each group was detected by Western-blot. Results Compared with the control group, the number of apoptotic cells in SH-SY5Y cells was significantly increased after treatment with 1 mmol / L MPP ~ + for 48 h. The effects of 1, 3 and 5 μmol / L SchA on the expression of MPP + Induced apoptosis of SH-SY5Y cells, and with the increase of SchA concentration, the number of apoptotic cells was significantly reduced. Western-blot results showed that cleaved caspase-3 protein expression in MPP ~ + group was significantly increased, while cleaved caspase-3 protein expression was significantly decreased in SchA intervention group (P <0.05). CONCLUSION: 1 mmol / L MPP ~ + can induce apoptosis in SH-SY5Y cells. However, certain concentrations of SchA can effectively inhibit the apoptosis of SH-SY5Y cells induced by MPP ~ +, and this effect may be through inhibiting the activation of caspase-3 protein Achieve.