丹皮酚抑制帕金森病模型细胞凋亡的作用

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...

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Published in浙江大学学报(医学版) no. 1; pp. 30 - 36
Main Author 王浩 耿赵铭 胡智伟 王舒燕 赵冰
Format Journal Article
LanguageChinese
Published 浙江省立同德医院神经内科,浙江杭州,310012%潍坊医学院临床医学系,山东潍坊,261053%浙江大学医学院附属第一医院麻醉科,浙江杭州,310003 2015
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ISSN1008-9292
DOI10.3785/j.issn.1008-9292.2015.01.005

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Summary: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。
Bibliography:33-1248/R
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
ISSN:1008-9292
DOI:10.3785/j.issn.1008-9292.2015.01.005