MPP^+诱导SK-N-SH细胞热休克蛋白的表达研究  

Expression of Heat Shock Proteins in SK-N-SH Cell Line Induced by MPP^+

作　　者：范国华[1] 陈生弟[1] 戚辰[1] 赵静[1] 

机构地区：[1]上海交通大学医学院瑞金医院神经科

出　　处：《上海交通大学学报（医学版）》2006年第2期130-134,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点基础研究发展规划("973"项目)(G1999054008);国家自然科学基金(39970263;30171025);上海市卫生系统百名跨世纪优秀学科带头人培养计划(97BR001)资助项目

摘　　要：目的研究SK-N-SH细胞在受到1-甲基-4-苯基-吡啶离子(MPP+)损伤时,细胞内热休克蛋白(HSPs)的表达。方法SK-N-SH细胞中加入不同浓度的MPP+制备帕金森病(PD)细胞模型,分别于培养24 h和48 h时,检测细胞存活率,选择合适的MPP+实验浓度。免疫印迹法(W estern b lot)和免疫细胞化学法观察细胞内Hsp70、Hsp40(HD J-1和HD J-2)和Hsp90的变化。结果细胞存活率检测表明MPP+合适的实验浓度为10~500μmol/L。W estern b lot和免疫细胞化学法检测均显示250μmol/L MPP+处理24 h,细胞内Hsp70含量明显升高(P<0.05);MPP+处理48 h,Hsp70含量随MPP+浓度的升高而下降;Hsp40/HD J-1和Hsp90的变化趋势基本与Hsp70相同;而HD J-2在细胞内的含量无明显改变(P>0.05)。结论HSPs的含量在MPP+制备的PD细胞模型中,随MPP+作用时间的延长而呈现先增加后降低的趋势。Objective To investigate the expression of heat shock proteins （HSPs） in the human neuroblastoma SK-N-SH cell line which was induced by 1-methyl-4-phenylpyridinium ion （ MPP ^＋ ）. Methods Cell viability was measured 24 and 48 h after incubation of various concentrations of MPP ^＋ in SK-N-SH cells. After appropriate concentration ranges of MPP ^＋ were determined, the expressions of Hsp70, Hsp40 （ HDJ-1 and HDJ-2） and Hsp90 in SK-N-SH cells were detected by Western blot and immunocytochemistry. Results 10-500 μomol/L MPP ^＋ were chosen for subsequent research. The expression of Hsp70 in SK-N-SH cells was significantly increased after being treated with 250 pomol/L MPP ^＋ for 24 h （ P 〈 0.05）. The expression of Hsp70 was decreased with the elevation of MPP ^＋ concentration after being treated with MPP ^＋ for 48 h. The expression trend of HDJ-1 and Hsp90 was quite similar to that of Hsp70. However, no significant alteration was found in the expression of HDJ-2 after MPP^＋ treated （P 〉0.05）. Conclusion The expressions of HSPs in the MPP ^＋-induced cell model of Parkinson＇s disease were elevated at first and then lowered down with time.

关 键 词：帕金森病 热休克蛋白 1-甲基-4-苯基-吡啶离子 MPP^+诱导 SK-N-SH细胞热休克蛋白 

分 类 号：R742.5[医药卫生—神经病学与精神病学] Q786[医药卫生—临床医学]