侧脑室注射DJ-1蛋白对MPTP致帕金森病小鼠的神经保护作用  

Neuroprotective effects of intracerebroventricular administration of DJ-1 protein on MPTP-induced Parkinson's disease in mice

作　　者：孙双勇[1,2] 周俊俊[1,2] 邓政[1,2] 安春娜[1,2] 赵欣[1,2] 蒲小平[1,2] 

机构地区：[1]北京大学药学院分子与细胞药理学系 [2]北京大学天然药物与仿生药物国家重点实验室,北京100191

出　　处：《中国新药杂志》2013年第3期339-344,共6页Chinese Journal of New Drugs

基　　金：教育部博士点基金资助项目(20090001110069)

摘　　要：目的:观察DJ-1蛋白对1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahy-dropyridine,MPTP)致帕金森病(Parkinson's disease,PD)小鼠是否具有神经保护作用。方法:分别采用SDS-PAGE和生物质谱仪对DJ-1蛋白的纯度和活性位点进行检测。然后将DJ-1蛋白注射进入小鼠侧脑室内,24 h后小鼠腹腔注射MPTP造模,通过滚筒实验、自主活动实验和竖直网格实验观察行为学变化,使用HPLC-ECD对纹状体内神经递质含量进行测定,通过酪氨酸羟化酶(tyroxine hydroxylase,TH)免疫组化染色观察SN区域多巴胺能神经元的存活状态。结果:SDS-PAGE和质谱实验表明DJ-1蛋白纯度较高,且活性位点未产生氧化;将DJ-1蛋白注射进入PD小鼠侧脑室后,小鼠行为学明显改善,纹状体内神经递质含量降低有所恢复,黑质区域多巴胺能神经元细胞死亡显著减少。结论:侧脑室注射DJ-1蛋白能够显著改善MPTP致的小鼠帕金森病症状,具有一定的神经保护作用。Objective： To observe the neuroprotective effects of D J-1 protein on 1-methyl-4- phenyl-1,2,3, 6-tetrahydropyridine （MPTP） induced Parkinson＇s disease （PD） in mice. Methods： SDS-PAGE and bio-mass spectrometry were used to determine the purity and active sites of DJ-1 protein. DJ-1 protein was then injected into the lateral ventricles of mice. MPTP was intraperitoneally injected to induce Parkinson＇s disease after 24 hours. Be- havioral changes of mice in each group were observed through rotarod performance test, spontaneous movement test and vertical grid test. Neurotransmitter contents in the striatum were determined by HPLC-ECD, and dopaminergic neurons in the Substantia nigra （SN） were observed through tyroxine hydroxylase （TH） immunohistoehemical stai- ning. Results： The results of SDS-PAGE and bio-mass spectrometry indicated that D J-1 protein had high purity and it was not oxidized. The PD symptoms of mice were improved obviously after D J-1 protein was injected into the lat- eral ventricles of mice, including behavioral improvement, neurotransmitter contents restoration and dopaminergic neuronal death reduction. Conclusion： Intracerebroventricular administration of D J-1 protein can improve PDsymptoms of mice, and D J-1 protein is proved to have neuroprotective effects.

关 键 词：DJ-1蛋白 帕金森病 1-甲基-4-苯基-1  2  3  6-四氢吡啶 神经递质 多巴胺能神经元 

分 类 号：R966[医药卫生—药理学]