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Journal of ZheJiang University(Medical Science)  2015, Vol. 44 Issue (1): 30-36    DOI: 10.3785/j.issn.1008-9292.2015.01.005
    
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
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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 words1-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     
Received: 16 May 2014     
CLC:  R96  
Cite this article:

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.

URL:

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


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、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 

[1] SCHAPIRA A H, JENNER P. Etiology and pathogenesis of Parkinson's disease [J]. Mov Disord, 2011, 26(6):1049-1055.
[2] PERIER C, BOVE J, VILA M. Mitochondria and programmed cell death in Parkinson's disease: apoptosis and beyond [J]. Antioxid Redox Signal, 2012, 116(9):883-895.
[3] BISAGLIA M, FILOGRANA R, BELTRAMINI M, et al. Are dopamine derivatives implicated in the pathogenesis of Parkinson's disease? [J]. Ageing Res Rev, 2014,13: 107-114.
[4] ESPOSITO E, CUZZOCREA S. New therapeutic strategy for Parkinson's and Alzheimer's disease [J]. Curr Med Chem, 2010, 17(25):2764-2774.
[5] 王 浩, 石巧娟, 史文珍, 等. 半胱氨酰白三烯受体CysLT_1R和CysLT_2R以及GPR17在帕金森病模型小鼠脑内的表达分布[J]. 浙江大学学报(医学版), 2013, 42(1):52-60. WANG Hao, SHI Qiao-juan, SHI Wen-zhen, et al. Expression and distribution of cysteinyl leukotriene receptors CysLT1R and CysLT2R, and GPR17 in brain of Parkinson disease model mice [J]. Journal of Zhejiang University(Medical Science), 2013,42(1):52-60. (in Chinese)
[6] 张晓燕, 张丽慧, 李成檀, 等. 5-脂氧酶参与鱼藤酮诱导的PC12细胞损伤[J]. 浙江大学学报(医学版), 2011,40(2):150-155. ZHANG Xiao-yan, ZHANG Li-hui, LI Cheng-tan, et al. 5-lipoxygenase is involved in rotenone-induced injury in PC12 cells [J]. Journal of Zhejiang University(Medical Science), 2011,40(2):150-155. (in Chinese)
[7] 郭 齐, 李贻奎, 王志国, 等. 丹皮酚药理研究进展[J]. 中医药信息, 2009,29(1):20-22. GUO Qi, LI Yi-kui, WANG Zhi-guo, et al. Research progress on paeonol pharmacological [J]. Information on Traditional Chinese Medicine, 2009, 29(1): 20-22. (in Chinese)
[8] ZHAO Y, FU B, ZHANG X, et al. Paeonol pretreatment attenuates cerebral ischemic injury via upregulating expression of pAkt, Nrf2, HO-1 andameliorating BBB permeability in mice [J]. Brain Res Bull, 2014, 109: 61-67.
[9] WU J B, SONG N N, WEI X B, et al. Protective effects of paeonol on cultured rat hippocampal neurons against oxygen-glucose deprivation-induced injury [J]. J Neurol Sci, 2008, 264 (1-2):50-55.
[10] ZHOU J, ZHOU L, HOU D, et al. Paeonol increases levels of cortical cytochrome oxidase and vascular actin and improves behavior in a rat model of Alzheimer's disease [J]. Brain Res, 2011, 1388: 141-147.
[11] ZHONG S Z, GE Q H, QU R, et al. Paeonol attenuates neurotoxicity and ameliorates cognitive impairment induced by d-galactose in ICR mice [J]. J Neurol Sci, 2009, 277(1-2):58-64.
[12] LIU J, FENG L, MA D, et al. Neuroprotective effect of paeonol on cognition deficits of diabetic encephalopathy in streptozotocin-induced diabetic rat [J]. Neurosci Lett, 2013, 549: 63-68.
[13] SU S Y, CHENG C Y, TSAI T H, et al. Paeonol protects memory after ischemic stroke via inhibiting β-secretase and apoptosis [J]. Evid Based Complement Alternat Med, 2012, 2012: 932823.
[14] 王 浩, 耿赵铭, 呙登俊, 等. 丹皮酚对血管性认知障碍小鼠认知功能的保护作用[J]. 温州医科大学学报, 2014,44(10):708-711. WANG Hao, GENG Zhao-ming, GUO Deng-jun, et al. Protective effects of paeonol on cognitive dysfunctions in mice with vascular cognitive impairment [J]. Journal of Wenzhou Medical University, 2014,44(10):708-711. (in Chinese)
[15] TSENG Y T, HSU Y Y, SHIH Y T, et al. Paeonol attenuates microglia-mediated inflammation and oxidative stress-induced neurotoxicity in rat primary microglia and cortical neurons [J]. Shock, 2012,7(3):312-318.

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