丹皮酚抑制帕金森病模型细胞凋亡的作用

Neuroprotective effects of paeonol in a cell model of Parkinson disease

目的：观察丹皮酚对帕金森病模型细胞凋亡的影响。方法：采用1－甲基－4－苯基吡啶（ MPP＋）处理具有多巴胺能神经元特性的PC12细胞建立帕金森病体外模型，并分为正常对照组、空白对照组、1μmol／L丹皮酚组、3μmol／L丹皮酚组和9μmol／L丹皮酚组。以四甲基偶氮唑蓝比色法和乳酸脱氢酶法检测细胞损伤，以赫斯特荧光染剂染色及流式细胞术检测细胞凋亡，以二氯二氢荧光素－乙酰乙酸酯法检测细胞活性氧生成，以蛋白质印迹法检测凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶－3（caspase－3）、Bcl－2和Bcl－2相关X蛋白（Bax）的表达水平。结果：与正常对照组比较，空白对照组细胞存活率显著降低，乳酸脱氢酶漏出率升高，凋亡细胞增多，活性氧生成增加，凋亡相关分子caspase－3活性上调、Bax／Bcl－2比值升高，差异均具有统计学意义（均P＜0．01）。与空白对照组比较，各浓度丹皮酚预处理后细胞存活率显著升高，乳酸脱氢酶漏出率降低，凋亡细胞减少，并抑制活性氧生成，降低Bax／Bcl－2的比值及caspase－3蛋白水平，差异均具有统计学意义（ P＜0．05或P＜0．01）。结论：丹皮酚能抑制帕金森病模型PC12细胞凋亡，其发挥保护作用的机制可能与改善氧化应激、降低Bax／Bcl－2比值、抑制caspase－3活化有关。

Objective：To investigate the effects of paeonol on neuron cell model of Parkinson disease （ PD） .Methods： The cell model of Parkinson disease was induced by treatment of 1-Methyl-4-phenylpyridinium （ MPP＋） in PC12 cells, the PD model cells were treated with 1 μmol/L, 3 μmol/L or 9 μmol/L paeonol for 24h, respectively.Cell viability and LDH leakage were detected by MTT and lactate dehydrogenase （ LDH） assay; the apoptosis of PC12 cells was assessed by Hoechst 33258 staining and flow cytometry; reactive oxygen species （ ROS ） production was detected by DCFH-DA method;and the ratio of Bax/Bcl-2 and activation of caspase-3 were determined by Western blotting .Results： MPP＋treatment significantly reduced cell viability , increased LDH leakage , enhanced the proportion of apoptotic cells and ROS production .In addition , MPP＋ treatment dramatically increased the Bax/Bcl-2 ratio, and the activation of caspase-3.Compared to PD model group , paeonol treatment significantly enhanced cell viability , decreased LDH leakage , inhibited the proportion of apoptotic cells and ROS production , reduced the Bax/Bcl-2 ratio and the activated caspase-3 protein .Conclusion：Paeonol can prevent PC 12 cells from apoptosis induced by MPP＋, and the mechanism may be associated with the down-regulation of ROS production, Bax/Bcl-2 ratio and Caspase-3 activation.