脑卒中前敲减HDAC2改善光照脑缺血模型小鼠的行为学缺陷
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国家自然科学基金重点项目(31530091);科技部973计划(2011CB504404)


HDAC2 knockdown before stroke improves behavioral deficits of photothrombotic stroke mice
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    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(2 μL)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。

    Abstract:

    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/flox mice,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.

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袁红瑾,林瑜辉,董 健,朱东亚.脑卒中前敲减HDAC2改善光照脑缺血模型小鼠的行为学缺陷[J].南京医科大学学报(自然科学版),2017,(9):1154-1158

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  • 收稿日期:2016-07-31
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  • 在线发布日期: 2017-09-25
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