过表达PINK1抵抗鱼藤酮引起多巴胺神经元损伤的研究  

Overexpression of PINK1 in C57BL/6 Mice Striatum May Alleviate Injury Caused by Rotenone

作　　者：龚普盛[1] 张建亮[1] 付越姣[1] 贾焕珍[1] 段春礼[1] 鲁玲玲[1] 赵春礼[1] 杨慧[1] 

机构地区：[1]首都医科大学神经科学研究所北京市神经再生修复研究重点实验室神经变性病教育部重点实验室北京100069

出　　处：《中国生物工程杂志》2012年第2期33-38,共6页China Biotechnology

基　　金：国家重点基础研究发展计划(2011CB504103);国家自然科学基金(30970940);北京市教育委员会科技发展计划重点项目(KZ201010025022);北京市自然基金面上项目(5102012);北京市高校人才强教资助PHR(200907113)资助项目

摘　　要：目的:明确在C57BL/6小鼠纹状体过表达野生型的人源帕金森相关蛋白PINK1能否减轻由侧脑室注射鱼藤酮引起多巴胺神经元损伤。方法:通过向C57BL/6小鼠(雄性,7周龄,18～20g)左侧纹状体中注射带有GFP人源野生型PINK1及突变体PINK1G309D的慢病毒包装颗粒,两周后向小鼠左侧侧脑室中定位注射鱼藤酮,通过蛋白质印迹,免疫组化和行为学的方法检测PINK1对鱼藤酮引起多巴胺神经元损伤的影响。结果:蛋白质印迹和免疫组化的实验都证明了在C57BL/6小鼠纹状体过表达野生型的PINK1对于鱼藤酮引起多巴胺能神经元的减少有明显的抑制作用(P<0.01),但对鱼藤酮引起的行为学损伤没有明显的改善作用。Background ： The etiology of Parkinson ＇ s disease （ PD ） involves genetic, environmental, endogenous neurotoxins and other factors. But the current weight of evidence strongly suggests that environmental neurotoxins may play an important role in the etiology of PD. Rotenone is a classical mitochondrial complex-I inhibitor and is the most potent member of the rotenoids, a family of isoflavonoids extracted from Leguminosae plants. More recently, chronic intraperitoneal injection of rotenone has been shown to cause dosedependent dopamine （DA） loss, reduced tyrosine hydroxylase （TH） immunoreactivity, and behavioral abnormalities in rats. WEN-induced kinase 1 （PINK1） is linked to recessive PD. Pink1 deletion results in impaired DA release and decreased mitochondrial respiration in the striatum of mice. Objective ： To test if overexpression of PINK1 in C57BL/6 mice striatum may alleviate the injury of dopaminergic neuron caused by Rotenone. Methods： Lentivirns of GFP-PINK1-LV, GFP-PINK1G3~gD-LV or GFP-vector-LV were injected into the C57BL/6 mice （male, 7 weeks old,18-20g weight） striatum. Two weeks later, Rotenone dissolved in DMSO was injected into the C57BL/6 mice lateral ventricles. After 2 weeks, TH protein level in C57BL/6 mice striatum was measured by Western blotting and Immunohistochemieal staining. Meanwhile the behavioral performance was evaluated in Open Filed experment. Results： Western blotting and immunohistochemical experiments have proved that over- expression of wild-type PINK1 in C57BL/6 mice striatum may rescue the TH protein level reduction, but can not alleviate the behavioral injury caused by Rotenone.

关 键 词：PINK1(PTEN-induced kinase 1) 鱼藤酮(Rotenone) 酪氨酸羟化酶(Tyrosine hydroxylase) 

分 类 号：Q819[生物学—生物工程]