电针激活大脑中动脉闭塞大鼠miRNA调控NF.pdf

ChineseJournalofRehabilitationMedicine, Apr.2016,Vol. 31, No.4 ·基础研究· 电针激活大脑中动脉闭塞大鼠miRNA调控 NF-κB信号通路的机制研究 姚建宁 王 鲜 柳维林 上官豪 郑 薏 黄 佳 陶 静 陈立典1 1 1 1 1 1 1 1,2 摘要 目的：探讨电针通过NF-κB 信号通路治疗局灶性脑缺血再灌注损伤大鼠所产生的抗炎作用的miRNA 调控机制。 方法：将54 只SD 大鼠随机分为假手术组、模型组及电针组，以大脑中动脉闭塞（middle cerebral artery occlusion, MCAO ）法建立大鼠局灶性脑缺血再灌注模型。利用TargetScan 预测miR-9a 的靶基因，并用双荧光素酶报告载体验 证。运用HE 染色及TTC 染色观察脑缺血后大鼠皮质炎症反应及梗死面积；应用蛋白免疫印迹法和实时PCR 检测 缺血侧皮质组织中NF-κB ，TNF-α及miR-9a 的表达情况。 结果：电针曲池、足三里穴可明显改善MCAO 大鼠的神经功能缺损症状，降低脑梗死范围，炎症反应明显缓解。可 以提高缺血侧皮质miR-9a 的表达（P ＜0.05 ），降低炎症因子NF-κB ，TNF-α的表达（P ＜0.05 ）。 结论：电针调控NF-κB 信号通路，抑制炎症因子分泌，来实现对脑缺血的治疗作用的机制可能与促进miR-9a 的表达 上调相关。 关键词 电针；脑缺血；微小RNA-9a ；NF-κB 信号通路 中图分类号：R743.3, R245 文献标识码：A 文章编号：1001-1242(2016)-04-0388-06 The anti-inflammatory function of miRNA regulation of NF-κB signaling pathway by Electroacupuncture on Middle cerebral artery occlusion rats/YAO Jianning, WANG Xian, LIU Weilin, et al.//Chinese Journal ofRehabilitationMedicine, 2016, 31(4): 388—393 Abstract Objective: To investigate the possible anti- inflammatory mechanism of electroacupuncture (EA) via regulating NF-κB pathway in rats with focal cerebral ischemia-reperfusion (I/R). Method: Fifty- four male adult SD rats were randomly divided into the sham operation control group (SC group), the ischemia model group (IC group) and the electroacupuncture group (EA group). Middle cerebral ar- tery occlusion (MCAO) was performed to establish the focal cerebral ischemia- reperfusion injury model. The potential target genes of miR- 9a was predicted and identified by dualluciferase reporter assay. TTC stain was used to examine the volume of infarction, HE staining was observed inflammatory responses. Western blot anal- ysis was employed to determine the expression of NF- κB