摘要
目的:探讨高迁移率族蛋白B1(HMGB1)对局灶脑缺血/再灌注大脑皮质梗死周围区神经干细胞增殖的影响。方法:选取雄性SD大鼠48只,分为假手术组、模型组、RNA干扰组和阴性干扰组。采用线栓法制备大鼠局灶脑缺血/再灌注模型,缺血1 h再灌注7 d时观测。用携带大鼠HMGB1 shRNA的慢病毒载体介导RNA干扰抑制脑内HMGB1的表达,通过免疫荧光双标记HMGB1/GFAP和Western blotting法检测RNA干扰效果。通过免疫荧光双标记5-溴脱氧尿嘧啶(BrdU)/神经巢蛋白(nestin)检测大脑皮质梗死周围区神经干细胞的增殖。结果:与假手术比较,再灌注7 d时,模型组大脑皮质梗死周围区HMGB1蛋白表达明显升高(P<0.05);与模型组比较,RNA干扰有效抑制HMGB1蛋白的表达(P<0.05)。与假手术组比较,模型组的BrdU/nestin双标记细胞明显增多(P<0.05);与模型组比较,RNA干扰组的BrdU/nestin双标记细胞明显减少(P<0.05)。结论:局灶脑缺血/再灌注后大脑皮质梗死周围区HMGB1蛋白表达水平升高可促进该部位的神经干细胞增殖。
AIM： To investigate the influence of high-mobility group box 1（ HMGB1） on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia /reperfusion model rats. METHODS： Male SD rats（ n =48） were randomly divided into sham group,ischemia/reperfusion（ I/R） group,RNA interference group and negative interference group. The rat middle cerebral artery was blocked to establish focal cerebral I /R model（ ischemia for 1 h and reperfusion for 7 d）. Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain. The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB1 /GFAP and Western blotting. The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labeling of BrdU /nestin. RESULTS： The protein expression of HMGB1 in I /R group was much higher than those in sham group（ P 〈0. 05）. RNA interference effectively inhibited the HMGB1 expression（ P 〈0. 05）. Double labeled BrdU /nestin positive cells in I /R group were more than that in sham group（ P〈 0. 05）. The double labeled BrdU /nestin positive cells were significantly decreased in RNA interference group（ P 〈0. 05）. CONCLUSION： Focal cerebral ischemia /reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB1 protein level.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第9期1616-1621,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30470606)
重庆市卫生局中医药科研项目(No.2003-B-16)
