论文部分内容阅读
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 investigate the role of pre-stroke histone deacetylase 2 (HDAC2) in behavioral deficits induced by light-induced thrombotic cerebral ischemia in mice. Methods: The adeno-associated virus AAV-CAG-EGFP-Cre (2μL) was injected into the motor cortex of HDAC2flox / flox mice to knock down HDAC2 by micro-drug delivery system. The virus AAV was confirmed by immunofluorescence and Western blot analysis -CAG-EGFP-Cre and control virus AAV-CAG-EGFP. 7 days after light-induced thrombotic cerebral ischemia modeling, and 3 days before modeling and 8 days after modeling using grid experiments and cylinder experiments were detected before and after modeling AAV-CAG-EGFP-Cre Behavioral changes of AAV-CAG-EGFP group. Results: Compared with control AAV-CAG-EGFP, AAV-CAG-EGFP-Cre could knockdown HDAC2 specifically, and immunofluorescence showed a decrease of double positive cells of GFP and HDAC2. Western blot showed that HDAC2 expression level Decreased (P <0.05). Behavioral testing before and after cerebral ischemia showed that downregulation of HDAC2 had no effect on pre-ischemic behavior, but significantly reduced the rate of loss of the grid after brain ischemia (P <0.001) and that of the cylinder experiment The Symmetry Index (P <0.001) promoted behavioral improvement. Conclusion: Knockdown of HDAC2 by recombinant virus before stroke can improve the behavioral deficits in mice after stroke.