DJ-1保护大鼠中脑黑质多巴胺能神经元抵抗鱼藤酮损伤的研究  

The Protective Effects of DJ-1 Against Rotenone Induced Dopaminergic Neuron Injury in the Rat SN

作　　者：隋雷鸣[1] 高华[2] 谷利[3] 杨巍巍[3] 杨慧[3] 

机构地区：[1]首都医科大学医学实验与测试中心,北京100069 [2]北京市神经外科研究所,北京100050 [3]首都医科大学神经生物学系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》2013年第5期7-12,共6页China Biotechnology

基　　金：国家“973”计划(2011CB504102,2012CB722407);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);北京市神经再生修复研究重点实验室开放课题(2011SJZS02);神经变性病教育部重点实验室开放课题(2011SJBX03)

摘　　要：目的:探讨过表达DJ-1蛋白能否保护大鼠中脑黑质多巴胺能神经元抵抗鱼藤酮所致的急性损伤。方法:构建表达DJ-1基因的腺相关病毒载体(rAAV-DJ-1)并转染HEK-293细胞,进行腺相关假病毒颗粒包装,所得假病毒颗粒注射到大鼠脑内感染黑质神经细胞;4周后,注射鱼藤酮于大鼠黑质相同脑区;通过免疫组织化学和免疫印迹试验鉴定TH蛋白表达情况,采用动物行为轨迹分析软件计算大鼠30 min内运动距离以及强迫游泳试验鉴定大鼠精神症状。结果:与对照组相比,过表达DJ-1组大鼠运动损伤症状较轻,精神症状改善明显;损伤侧黑质TH阳性神经元数目和TH蛋白表达水平显著高于对照组。结论:过表达DJ-1蛋白能保护大鼠黑质多巴胺能神经元抵抗鱼藤酮所致的急性损伤,延缓多巴胺能神经元的退行性变,提示DJ-1可能对帕金森病的治疗有较好的应用前景。Objective： To observe protective effects of DJ-1 against rotenone induced dopaminergic neuron injury in rat substantia nigra （SN）. Methods： DJ-1 gene was reconstructed into adeno-associated virus （AAV） vector. Then rAAV-DJ-1 was transfected into HEK-293 cells for packaging. AAV vectors encoding human DJ-1protein were stereotactically injected into the rat SN 4 weeks prior to rotenone lesion in SN. Tyrosine hydroxylase （TH） protein was determined using immunohistochemical staining and Western blot. Ethovison software allowed measurement of movement distance in 30 minutes. Rotenone-induced depression-like behaviors in rats was measured by swimming. Results： The animals of the AAV-DJ-1 group significantly increased movement distance and swimming time comparing to control one. Furthermore, overexpression of DJ-1 dramatically inhibited the loss of TH-positive neurons. Conclusion ： Overexpression of D J-1 protected the dopaminergic neurons against rotenone induced injury in the rat SN, and strongly support the potential of DJ-1 gene on drug therapy for PD.

关 键 词：DJ-1 鱼藤酮 酪氨酸羟化酶 帕金森病 

分 类 号：R741[医药卫生—神经病学与精神病学]