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目的探讨Cav1/Notch1/Hes1通路在小鼠脑缺血后海马神经干细胞(neural stem cells,NSCs)增殖中的作用及补阳还五汤促神经再生的机制。方法采用大脑中动脉阻塞法(middle cerebral artery occlusion,MCAO)复制小鼠脑缺血模型,将野生型(wild type,WT)和小凹蛋白-1基因敲除小鼠(caveolin1 gene knockout,Cav1 KO)分别随机分为假手术组、模型组、补阳组,每组12只,腹腔注射Brd U标记增殖细胞,干预7 d后,采用免疫荧光Brd U/Nestin双标法检测小鼠缺血侧海马NSCs增殖情况,Western blot法检测Notch1、NICD、Hes1蛋白表达,Real time PCR法检测Notch1、Hes1m RNA表达。结果 MCAO法成功复制了小鼠脑缺血模型;WT模型组、补阳组和KO模型组、补阳组缺血侧海马Brd U/Nestin双标阳性细胞,Notch1、NICD、Hes1蛋白表达及Notch1、Hes1m RNA表达均增加,与相应假手术组比较,差异有统计学意义(P<0.01,P<0.05),同时,WT模型组要高于KO模型组(P<0.05),WT补阳组要高于WT模型组和KO补阳组(P<0.01,P<0.05)。结论Cav1/Notch1/Hes1通路在小鼠脑缺血后海马NSCs增殖中发挥重要调控作用;通过上调Cav1/Notch1/Hes1通路活性是补阳还五汤促进缺血海马神经再生作用机理之一。
Objective To investigate the role of Cav1 / Notch1 / Hes1 pathway in the proliferation of neural stem cells (NSCs) after cerebral ischemia in mice and the mechanism of Buyang Huanwu Decoction promoting nerve regeneration. Methods Mice model of cerebral ischemia was induced by middle cerebral artery occlusion (MCAO). Wild type (WT) and caveolin1 gene knockout (Cav1 KO ) Were randomly divided into sham-operation group, model group and buxu-yang group, 12 rats in each group were injected intraperitoneally with Brd U labeled proliferating cells. After 7 days of intervention, immunofluorescence BrdU / Nestin double- The proliferation of hippocampal NSCs was detected by western blot. The expressions of Notch1, NICD and Hes1 were detected by Real-time PCR. Results The cerebral ischemia model was successfully replicated in MCAO mice. The expression of Notch1, NICD and Hes1 and the expression of BrdU / Nestin positive cells in hippocampus of WT, BuYang and KO model groups and Notch1 (P <0.01, P <0.05). Meanwhile, WT model group was higher than KO model group (P <0.05), WT supplementing group Higher than WT model group and KO supplement group (P <0.01, P <0.05). Conclusions The Cav1 / Notch1 / Hes1 pathway plays an important regulatory role in the proliferation of hippocampal NSCs after cerebral ischemia in mice. One of the mechanisms of Buyang Huanwu Decoction can promote neural regeneration of hippocampus by up-regulating the activity of Cav1 / Notch1 / Hes1 pathway.