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浙江大学学报（医学版）
浙江大学学报（医学版）  2015, Vol. 44 Issue (1): 30-36    DOI: 10.3785/j.issn.1008-9292.2015.01.005
专题报道     
丹皮酚抑制帕金森病模型细胞凋亡的作用
王浩1, 耿赵铭2, 胡智伟1, 王舒燕3, 赵冰3
1. 浙江省立同德医院神经内科, 浙江 杭州 310012;
2. 潍坊医学院临床医学系, 山东 潍坊 261053;
3. 浙江大学医学院附属第一医院麻醉科, 浙江 杭州 310003
Neuroprotective effects of paeonol in a cell model of Parkinson disease
WANG Hao1, GENG Zhao-ming2, HU Zhi-wei1, WANG Shu-yan3, ZHAO Bing3
1. Department of Neurology, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China;
2. Department of Clinical Medicine, Weifang Medical College, Weifang 261053, China;
3. Department of Anesthesiology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China
 全文: PDF(2362 KB)  
摘要：

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、1 μmol/L丹皮酚组、3 μmol/L丹皮酚组和9 μmol/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.
关键词： 1-甲基-4-苯基吡啶;  帕金森病;  细胞凋亡/药物作用;  牡丹酚/药理学;  牡丹酚/投药和剂量;  半胱氨酸天冬氨酸蛋白酶3/药物作用;  bcl-2相关X蛋白质;  基因, bcl-2;  活性氧    
Abstract:

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 PC12 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.
Key words: 1-Methyl-4-phenylpyridinium    Parkinson disease    Apoptosis/drug effect    Paeonol/pharmacology    Paeonol/administration &    dosage    Caspase 3/drug effects    bcl-2-associated X protein    Genes, bcl-2    Reactive oxygen species
收稿日期: 2014-05-16
CLC:  R96  
基金资助:

国家自然科学基金(81301122);高等学校博士学科点专项科研基金(20130101120029).
通讯作者: 赵 冰(1982-),女,博士,医师,从事神经药理学研究;E-mail: zhaobingzju@163.com     E-mail: zhaobingzju@163.com
作者简介: 王 浩(1982-),男,硕士,主治医师,从事帕金森病发病机制研究;E-mail: wangh8234@163.com
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引用本文:

王浩, 耿赵铭, 胡智伟, 王舒燕, 赵冰. 丹皮酚抑制帕金森病模型细胞凋亡的作用[J]. 浙江大学学报（医学版）, 2015, 44(1): 30-36.

WANG Hao, GENG Zhao-ming, HU Zhi-wei, WANG Shu-yan, ZHAO Bing. Neuroprotective effects of paeonol in a cell model of Parkinson disease. Journal of ZheJiang University(Medical Science), 2015, 44(1): 30-36.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2015.01.005    或    http://www.zjujournals.com/xueshu/med/CN/Y2015/V44/I1/30

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