脑卒中前敲减HDAC2改善光照脑缺血模型小鼠的行为学缺陷  

HDAC2 knockdown before stroke improves behavioral deficits of photothrombotic stroke mice

作　　者：袁红瑾 林瑜辉 董健 朱东亚[1] 

机构地区：[1]南京医科大学药学院药理实验室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2017年第9期1154-1158,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金重点项目(31530091);科技部973计划(2011CB504404)

摘　　要：目的:探讨脑卒中前蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)对于光诱导血栓形成性脑缺血模型小鼠行为学缺陷的作用。方法:通过微量给药系统将腺相关病毒AAV-CAG-EGFP-Cre(2μL)注射至HDAC2flox/flox鼠的运动皮层用以敲减HDAC2,通过免疫荧光以及免疫印迹(Western blot)分析确证病毒AAV-CAG-EGFP-Cre与对照病毒AAV-CAG-EGFP的表达。7 d后进行光诱导血栓形成性脑缺血造模,并于造模前第3天以及造模后第8天采用网格实验和圆筒实验分别检测造模前后AAV-CAG-EGFP-Cre组与AAV-CAG-EGFP组行为学的改变。结果:病毒AAV-CAG-EGFP-Cre相较于对照AAV-CAG-EGFP能特异性地敲减HDAC2,免疫荧光表现为GFP与HDAC2双阳性细胞数减少,Western blot显示针孔周围区HDAC2表达水平降低(P<0.05)。脑缺血前后的行为学检测表明病毒干预后,HDAC2的下调对缺血前行为学无影响,但显著降低了脑缺血后网格实验的失足率(P<0.001)与圆筒实验的不对称指数(P<0.001),促进了行为学改善。结论:脑卒中前通过重组病毒敲减HDAC2,可以改善脑卒中后小鼠行为学缺陷。Objective：To explore the influence of intervention of histone deacetylase 2 （HDAC2） on behavioristics of mice after photothrombotic stroke.Methods：Adeno-associated virus AAV-CAG-EGFP-Cre （2 μL） was microinjected through microcannula to the motor cortex of HDAC2flox/floxmice,aiming to knockdown HDAC2.And then,the expression of virus AAV-CAG-EGFP-Cre and its control AAV-CAG-EGFP was validated through immunofluorescence and Western blot.Seven days later,photothrombotic stroke was induced in the mice,and grid-walking task and cylinder task were conducted at day 3 before stroke and day 8 after stroke to detect behavior changes.Results：Compared with AAV-CAG-EGFP,AAV-CAG-EGFP-Cre significantly down-regulated HDAC2 expression,which was showed in the reduction of the number of GFP＋/HDAC2＋cells and the expression level of HDAC2 in the peri-pinhole zone （P〈0.05）.Behavioral tests demonstrated that the decrease of HDAC2 didn＇t influence the behavior before ischemia,but significantly reduced foot faults in the grid-walking task （P〈0.001） and asymmetry index in the cylinder task （P〈0.001） after ischemia,which improved behavioral deficits.Conclusion：HDAC2 knock down by viral interference can promote functional recovery after photothrombotic stroke.

关 键 词：组蛋白去乙酰化酶2 脑卒中 功能恢复 

分 类 号：R743.3[医药卫生—神经病学与精神病学]