小鼠缺血性脑损伤模型急性期海马SIRT3与自噬相关蛋白表达水平的相关性分析  

Correlative Analysis of Hippocampal SIRT3 with Autophagy-related Protein Expression in the Acute Period of Mouse Model of Cerebral Ischemia

作　　者：李建荣 张磊[1] 岳康异 罗鹏[1] 费舟[1] 蒋晓帆[1] LI Jian-rong;ZHANG Lei;YUE Kang-yi;LUO Peng;FEI Zhou;JIANG Xiao-fan(Department of Neurosurgery,Xijing Hospital,Fourth Military Medical University,Xi'an,Shaanxi,710032,China)

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032

出　　处：《现代生物医学进展》2018年第23期4401-4404,4408,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(8167050366)

摘　　要：目的:观察小鼠缺血性脑损伤模型急性期海马区域沉默信息因子3(SIRT3)和自噬相关蛋白的表达,并探讨两者的相关性。方法:选择C57BL/6J小鼠,采用大脑中动脉阻塞(MCAO)法建立缺血性脑损伤模型,并将小鼠随机分为假手术组(sham)和模型组(MCAO)。缺血性脑损伤急性期(6h),采用间接免疫荧光法观察小鼠海马CA1、CA3和DG区域SIRT3的表达,应用蛋白印迹法检测SIRT3、自噬相关蛋白LC3 I/II和Beclin-1的表达,而后用Spearman相关性分析明确SIRT3和LC3-II、Beclin-1表达的相关性。结果:海马各区域SIRT3阳性细胞数量在损伤后明显增多(P<0.05),且SIRT3蛋白表达也相对上调(P<0.05);损伤后自噬相关蛋白LC3-II和Beclin-1表达亦增高(P<0.05);Spearman相关性分析发现SIRT3与自噬相关蛋白LC3-II、Beclin-1表达均呈显著正相关(P<0.05)。结论:小鼠缺血性脑损伤模型急性期海马区域SIRT3和自噬相关蛋白的表达具有显著相关性,SIRT3对海马区域自噬的调节可能有重要作用。Objective: To explore the expression and correlation of hippocampal NAD dependent deacetylase sirtuin-3(SIRT3)with autophagy-related proteins in the acute period in a mouse model of cerebral ischemia. Methods: Cerebral ischemia was established using middle cerebral artery occlusion(MCAO) in C57BL/6J mice and randomly divided into the sham group and the MCAO group. In the acute period(6 h) of cerebral ischemia, immunofluorescence was used to observe the changes of SIRT3 expression in hippocampal CA1, CA3 and DG areas, and western blotting was used to analyze the relative protein expression of SIRT3 and autophagy-related proteins LC3 I/II and Beclin-1. Spearman correlative analysis was used to validate the relationship between SIRT3 and autophagy-related proteins expression. Results: SIRT3-positive cells were increased in the hippocampal subareas of MCAO groups compared with that of the sham group(P<0.05), and so do the protein levels of SIRT3(P<0.05);the protein expressions of LC3-II and Beclin-1 were also up-regulated in the MCAO group(P<0.05);Spearman correlational analysis presented the expression of SIRT3 and autophagy-related proteins(LC3-II and Beclin-1) were positively correlated(P<0.05). Conclusion: The expression of hippocampal SIRT3 and autophagy-related proteins were significantly correlated, indicating the essential role of SIRT3 in autophagy regulation.

关 键 词：沉默信息因子3 自噬 缺血性脑损伤 海马 

分 类 号：R-33[医药卫生] R651.15