自噬参与DJ-1保护大鼠黑质多巴胺能神经元抵抗鱼藤酮损伤

doi:10.3969/j.issn.1006-7795.2013.03.013

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (3): 385-390.doi: 10.3969/j.issn.1006-7795.2013.03.013

自噬参与DJ-1保护大鼠黑质多巴胺能神经元抵抗鱼藤酮损伤

刘慧丰¹, 杨巍巍², 高华³, 杨慧²

1. 北京王府中西医结合医院综合内科, 北京 100049;
2. 首都医科大学基础医学院神经生物学系北京脑重大疾病研究院, 北京 100069;
3. 首都医科大学附属北京天坛医院北京神经外科研究所,北京 100050

收稿日期:2013-04-08 出版日期:2013-06-21 发布日期:2013-06-17
通讯作者: 杨慧 E-mail:huiyang0105@sina.com
基金资助:
国家重点基础研究发展计划(2011CB504102,2012CB722407);国家自然科学基金(30970940);北京市自然科学基金重点项目(7131001)。

Autophagy is involved in DJ-1 protecting against rotenone-induced dopaminergic neurons injuries of substantia nigra in rats

LIU Huifeng¹, YANG Weiwei², GAO Hua³, YANG Hui²

1. Department of Internal Medicine, Beijing Wangfu Integrative Medicine Hospital, Beijing 100049;
2. Department of Neurobiology, School of Basic Medical Science, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069;
3. Beijing Tiantan Hospital, Beijing Neurosurgical Institute, Capital Medical University, Beijing 100050, China

Received:2013-04-08 Online:2013-06-21 Published:2013-06-17
Supported by:
This study was supported by National Basic Research Program of China (2011CB504102,2012CB722407), National Natural Science Foundation of China (30970940), Natural Science Foundation of Beijing (7131001).

摘要/Abstract

摘要：

目的观察过表达DJ-1是否缓解鱼藤酮致SD大鼠中脑黑质多巴胺能神经元损伤及其机制。方法在大鼠黑质致密部注射携带DJ-1、LacZ或者GFP-DJ-1、GFP的腺相关病毒颗粒,4周后注射鱼藤酮。通过免疫组织化学方法鉴定基因表达情况,并检测 TH阳性神经元数量,验证DJ-1保护作用。通过Western blotting方法检测自噬相关蛋白表达水平。结果鱼藤酮损伤4周时,可见过表达DJ-1组大鼠损伤侧黑质的TH阳性神经元数目显著高于单纯鱼藤酮损伤组,差异有统计学意义(P<0.05)。蛋白印迹检测动物损伤侧黑质自噬相关蛋白Beclin1、p-mTOR的表达和LC3 Ⅱ/Ⅰ的比值,发现DJ-1过表达组中Beclin1与LC3 Ⅱ/Ⅰ的比值均较对照组增加,差异有统计学意义;而p-mTOR的表达较对照组降低,差异有统计学意义(P<0.05)。结论 DJ-1能抵抗鱼藤酮对大鼠黑质多巴胺能神经元的损伤,DJ-1能上调自噬水平。

关键词: DJ-1, 鱼藤酮, 自噬, 大鼠

Abstract:

Objective To observe whether DJ-1 overexpression could attenuate rotenone-induced dopaminergic neurons injuries in substantia nigra (SN) of rats and its mechanism. Methods AAV vectors encoding human wild-type DJ-1, LacZ or GFP-DJ-1, GFP genes were stereotactically injected into the rat SN 4 weeks prior to rotenone lesion in SN. As identification of DJ-1 gene expression by immunohistochemistry, the numbers of TH positive neurons were detected to verify the neuron-protective effect of DJ-1. Autophagy-related proteins expression were investigated via Western blotting. Results After 4 weeks rotenone injected, the numbers of TH positive neurons increased significantly in impaired SN of rats in DJ-1 overexpressing group compared to rotenone-injected rats (P<0.05). Autophagy-related protein Beclin1, p-mTOR expression and LC3Ⅱ/Ⅰ ratio were detected in impaired SN of animals according to Western blotting. In DJ-1 overexpression group, Beclin 1 expression and the ratio of LC3Ⅱ to LC3I increased significantly compared with control groups, while p-mTOR protein expression decreased significantly compared with control groups (P<0.05).Conclusion DJ-1 could attenuate dopaminergic cells injuries induced by rotenone and upregulated autophagy level.

Key words: DJ-1, rotenone, autophagy, rat

中图分类号:

R 742.5

刘慧丰, 杨巍巍, 高华, 杨慧. 自噬参与DJ-1保护大鼠黑质多巴胺能神经元抵抗鱼藤酮损伤[J]. 首都医科大学学报, 2013, 34(3): 385-390.

LIU Huifeng, YANG Weiwei, GAO Hua, YANG Hui. Autophagy is involved in DJ-1 protecting against rotenone-induced dopaminergic neurons injuries of substantia nigra in rats[J]. Journal of Capital Medical University, 2013, 34(3): 385-390.

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自噬参与DJ-1保护大鼠黑质多巴胺能神经元抵抗鱼藤酮损伤

Autophagy is involved in DJ-1 protecting against rotenone-induced dopaminergic neurons injuries of substantia nigra in rats

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