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淫羊藿苷减轻MPP+对MES23.5细胞的神经毒性损伤

徐爱丽, 姜明春, 陈筱涵, 陈文芳

青岛大学医学院生理学教研室，山东省神经退变疾病协同创新中心，青岛 266071；滨州医学院生理学教研室，烟台 264003；泰山医学院生理学教研室，泰安 271016

摘要

淫羊藿苷是传统中药淫羊藿的主要活性成分。研究显示淫羊藿苷可能是一个潜在的抗老年疾病的药物。本研究旨在探讨淫羊藿苷对MPP+诱导的MES23.5细胞损伤的保护作用及其可能机制。细胞活力检测结果显示MPP+可剂量依赖性地损伤MES23.5细胞，降低细胞存活率，淫羊藿苷可以显著抑制MPP+所诱导的细胞毒性作用。此外，免疫细胞化学和免疫印迹结果显示淫羊藿苷可对抗MPP+引起的酪氨酸羟化酶(TH)阳性细胞数的减少和蛋白水平的降低。流式细胞术结果显示，淫羊藿苷可以逆转MPP+所导致的线粒体膜电位的降低。Real-time RT-PCR和Western blot结果显示，淫羊藿苷可逆转MPP+所导致的Bax基因和蛋白表达水平的上调以及Bcl-2基因和蛋白表达水平的下调；同时，淫羊藿苷也可以抑制MPP+所诱导的cleaved caspase-3蛋白的表达水平。以上结果提示，淫羊藿苷可明显对抗MPP+对MES23.5细胞的神经毒性作用，其作用机制可能与抗凋亡有关。

关键词： 淫羊藿苷; 1-甲基-4-苯吡啶离子; MES23.5细胞

分类号：R338

Icariin protects against MPP+-induced neurotoxicity in MES23.5 cells

XU Ai-Li, JIANG Ming-Chun, CHEN Xiao-Han, CHEN Wen-Fang

Department of Physiology, Shandong Provincial Collaborative Innovation Center for Neurodegenerative Disorders, Medical College of Qingdao University, Qingdao 266071, China； Department of Physiology, Binzhou Medical University, Yantai 264003, China； Department of Physiology, Medical College of Taishan, Taian 271016, China

Abstract

Icariin is the major bioactive component of Epimedium and has been demonstrated to be a potential drug for age-related diseases. The present study was aimed to investigate the neuroprotective properties of icariin against 1-methyl-4-phenylpyridinium ion (MPP+)-induced neurotoxicity in MES23.5 cells and the possible mechanisms. MTT assay showed that treatment with MPP+ attenuated the cell viability in a dose-dependent manner in MES23.5 cells. Icariin pretreatment resulted in an enhancement of survival. Immunocytochemistry analysis revealed that icariin treatment attenuated MPP+-induced loss of tyrosine hydroxylase (TH) positive cells. Meanwhile, Western blot confirmed MPP+ significantly decreased the TH protein expression, and icariin pretreatment could reverse the toxic effect of MPP+. Moreover, flow cytometry showed that MPP+-induced decrease of the mitochondrial membrane potential could be partly restored by icariin. Furthermore, real-time RT-PCR and Western blot analysis demonstrated that icariin treatment restored the MPP+-induced up-regulation of Bax and down-regulation of Bcl-2 mRNA and protein expressions. Western blot data also revealed the inhibitory effect of icariin on MPP+-induced up-regulation of cleaved caspase-3. These findings provide the evidence that icariin has neuroprotective properties against MPP+-induced neurotoxicity in MES23.5 cells and the mechanism might be related to the anti-apoptotic action of icariin.

Key words: icariin; 1-methyl-4-phenylpyridinium ion; MES23.5 cells

收稿日期：2016-02-25　　录用日期：2016-07-20

通讯作者：陈文芳　　E-mail: chenwenfangqd@163.com

引用本文：

徐爱丽, 姜明春, 陈筱涵, 陈文芳. 淫羊藿苷减轻MPP+对MES23.5细胞的神经毒性损伤[J]. 生理学报 2016; 68 (5): 585-591.

XU Ai-Li, JIANG Ming-Chun, CHEN Xiao-Han, CHEN Wen-Fang. Icariin protects against MPP+-induced neurotoxicity in MES23.5 cells. Acta Physiol Sin 2016; 68 (5): 585-591 (in Chinese with English abstract).