鱼藤酮对PC12细胞毒性作用的机制

doi:吉林省科技厅基金资助课题(200505200)

鱼藤酮对PC12细胞毒性作用的机制

韩威, 宫萍, 常明, 张瑜, 张颖, 王秋艳, 胡轶虹,胡林森

吉林大学第一医院神经内科，吉林长春 130021

收稿日期:2006-06-30 修回日期:1900-01-01 出版日期:2007-01-28 发布日期:2007-01-28
通讯作者: 胡林森

Mechanism of cytotoxicity induced by rotenone in PC12 cells

HAN Wei, GONG Ping, CHANG Ming , ZHANG Yu, ZHANG Ying, WANG Qiu-yan, HU Yi-hong , HU Lin-sen

Department of Neurology，First Hospital , Jilin University，Changchun 130021，China

Received:2006-06-30 Revised:1900-01-01 Online:2007-01-28 Published:2007-01-28
Contact: HU Lin-sen

摘要/Abstract

摘要： 目的：探讨鱼藤酮诱导PC12细胞毒性作用的可能机制，为神经防护药物的研发提供理论基础。方法：将培养的PC12细胞分为正常对照组和0.5μmol•L^-1鱼藤酮实验组，持续作用72 h，MTT法检测细胞存活率；Hoechst 33342检测细胞凋亡；提取实验组和对照组细胞的总蛋白，应用荧光差异凝胶电泳系统（DIGE）获得差异蛋白点的表达信息；通过MALDI-TOF 质谱鉴定差异蛋白点。结果：MTT法显示鱼藤酮实验组较正常对照组细胞存活率明显降低（P<0.01）；Hoechst 33342荧光染色显示鱼藤酮实验组可见较多凋亡细胞，表现为核边集、核固缩、核碎裂等特征性凋亡改变，其凋亡率明显高于对照组（P<0.01）。DIGE分析软件提示实验组点298的表达量高于对照组（P=0.004），点447的表达量低于对照组（P=0.009 5）。通过质谱分析和数据库检索，鉴定为纽蛋白和线粒体顺乌头酸酶。结论：纽蛋白和线粒体顺乌头酸酶参与细胞损伤，可能成为神经防护药物作用的靶点。

关键词: 鱼藤酮, 蛋白质组学, 细胞毒性作用

Abstract: To explore the mechanism of cytotoxity induced by rotenone in PC12 cells in order to provide the theoretical basis of neuroprotective drugs. Methods he cultivated PC12 cells were divided into control and 0.5　μmol•L^-1 rotenone-treated groups. Cell viability was estimated by MTT 72 h after treatment. Cell apoptosis was examined by Hoechst 33342. Proteins were extracted from two cell groups, respectively. The maps of proteins were set up by DIGE system. The altered protein spots were identified with MALDI-TOF MS and database searching. Results TT showed cell viability decreased significantly in 0.5 μmol•L^-1rotenone-treated group compared with control group（P<0.01）. Hoechst 33342 fluorescence staining showed that more apoptotic cells were found in rotenone-treated group, the ratio of apoptosis increased significantly compared with control group（P<0.01）. Cells showed condensed, fragmented and nuclei demonstrated by fluorescent dye, which were the characteristics of apoptosis. DIGE revealed two protein spots were significantly changed in rotenone-treated cells compared with control（P<0.01）. MALDI-TOF MS analysis and NCBI database searching demonstrated them as vinculin and mitochondrial aconitase, respectively. Conclusion Vinculin and mitochondrial aconitase may be involved in the cytotoxicity，which may be the target of neuroprotective drugs.

Key words: rotenone, proteomics, cytotoxicity

中图分类号:

R742.5

韩威, 宫萍, 常明, 张瑜, 张颖, 王秋艳, 胡轶虹,胡林森. 鱼藤酮对PC12细胞毒性作用的机制[J]. J4, 2007, 33(1): 98-101.

HAN Wei, GONG Ping, CHANG Ming , ZHANG Yu, ZHANG Ying, WANG Qiu-yan, HU Yi-hong , HU Lin-sen. Mechanism of cytotoxicity induced by rotenone in PC12 cells[J]. J4, 2007, 33(1): 98-101.

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