益气活血方基于miR-124调控Wnt通路促进神经再生-基础研究.PDFVIP

J South Med Univ, 2017, 37(8): 1047-1053 doi 10.3969/j.issn. 1673-4254.2017.08.08 ·1047 · 基础研究 益气活血方益气活血方基于基于miR-miR-124124调控调控WntWnt 通路促进通路促进神经再生神经再生 谭 辉，王 键，尹婷婷，何 玲，邓 勇，李 凤，王玉凤 安徽中医药大学新安医学教育部重点实验室，安徽 合肥 230038 摘要：目的探讨益气活血方（脑络欣通）对MCAO-R 气虚血瘀证大鼠神经再生的影响及可能的作用机制。方法 随机数字表法 将大鼠分成4 组：正常组、模型组、脑络欣通组、通心络组，其中脑络欣通组和通心络组分别用脑络欣通溶液、通心络溶液灌胃。 观察各组大鼠生命状态并进行神经功能缺损、气虚证和血瘀证评分，运用激光多普勒仪动态监测局部脑血流量（rCBF ），TTC 染 色检测脑梗死体积，免疫组化结合免疫荧光染色检测脑组织Nestin 和BrdU 表达，荧光定量PCR 检测miRNA- 124水平，Western blotting 检测Wnt3a、GSK3β和β-catenin 蛋白表达水平。结果 脑络欣通和通心络都能改善模型大鼠神经功能，降低气虚证和血 瘀证评分，减少脑梗死体积，rCBF 提高（P0.01）；脑络欣通和通心络组BrdU 表达增强，与正常组比较，其他3组阳性细胞数明显 增多（P0.01），与模型组比较，脑络欣通和通心络组显著增多，Nestin 表达明显增强（P0.01）。脑络欣通能抑制模型大鼠脑组织 miRNA- 124和Wnt3a 应激性的高表达，同时提高β-catenin 的表达水平（P0.01）。但脑络欣通和通心络对模型大鼠GSK3β蛋白 表达均无明显影响（P0.05）。结论 脑络欣通可能通过影响脑组织miRNA- 124 的表达而激活Wnt 通路，从而发挥对气虚血瘀 型脑缺血的神经保护以及神经再生的作用。 关键词：气虚血瘀证；脑缺血；脑络欣通；神经再生；miRNA- 124；Wnt 通路 Yiqihuoxue prescription promotes nerve regeneration by miR-124-mediated regulation of Wnt signaling in rats TAN Hui, WANG Jian,YIN Tingting, HE Ling, DENG Yong, LI Feng, WANG Yufeng Key Laboratory for Xin an Medicine of the Education Ministry, Anhui Chinese Medical University, Hefei 230038, China Abstract: Objective To investigate the effect of Yiqihuoxue prescription (NLXT) on nerve regeneration in MCAO-R rat models of qi deficiency and blood stasis syndrome and explore the underlying mechanisms. Methods The rats were randomized into 4 groups, namely the control group, model group, NLXT group and TXL group. The rats in NLXT group and TXL group were treated with gavage of NLXT and TXL solutions, respectively. The NFDS, QDSS and BSSS of the rats were evaluated. The regional cerebral blood flow (rCBF) were dynamically monitored with laser Doppler scanning, and the volume of cerebral infarction was detected with TTC-dye; the expression levels of nes