脑疏宁对缺血性脑卒中神经再生作用的研究

Effect of Nao-shuning on Nerve Regeneration in Ischemic Stroke

中药脑疏宁是基于中风本虚标实、气血亏虚、痰瘀闭阻的病因确立的活血、利水、通腑、调气的临床经验方,已有研究表明其在大鼠MCAO模型和TBI模型中具有改善脑损伤的作用,但在体外细胞实验上鲜有研究。应用体外分离培养大鼠神经干细胞,探讨脑疏宁对神经干细胞增殖的影响及其相关作用机制。本文采用WST-1法测定经不同浓度(21.4 mg/mL、2.14 mg/mL、214μg/mL、21.4μg/mL、2.14μg/mL、0)脑疏宁分别处理神经干细胞24 h和48 h后,其对细胞增殖的影响;利用U0126建立MAPK/ERK信号通路阻断模型,WST-1法检测脑疏宁对该阻断模型的作用;Western blot法检测脑疏宁处理后细胞中p-ERK1/2蛋白的表达情况。结果表明,与对照组相比,一定浓度范围内的脑疏宁可以显著促进神经干细胞的增殖(P<0.05),与浓度呈依赖关系,但高浓度脑疏宁抑制干细胞增殖并使细胞死亡。WST-1结果显示,脑疏宁可以使MAPK/ERK信号通路阻断模型组的干细胞增殖,但其增殖结果不显著。Western blot结果显示,脑疏宁能显著上调MAPK/ERK信号通路中p-ERK1/2的表达,促进ERK1/2磷酸化。适宜浓度的中药脑疏宁可以促进体外培养的神经干细胞增殖,其机制可能与促进MAPK/ERK信号通路中ERK1/2的磷酸化有关。

Nao-shuning is a Chinese compound prescription used in clinical experience. Many studies have shown that Nao-shuning can improve brain injury in rat MCAO model and TBI model,but few studies have been done on cell experiments. It is studied to explore the effects of Nao-shuning on neural stem cell proliferation and related mechanisms by culturing rat neural stem cells. Method: The WST-1 method was used to measure and observe the effect of the drug on cell proliferation after treating neural stem cells for 24 hours and 48 hours with different concentrations of Nao-shuning( 21. 4 mg/m L、2. 14 mg/m L、214 μg/m L、21. 4 μg/m L、2. 14 μg/m L、0) respectively. U0126 was used to establish the MAPK/ERK signaling pathway blocking model,and the effect of Nao-shuning on this blocking model was detected by WST-1 method. Western blot was used for detection of pERK1/2 protein expression in cells after Nao-shuning treatment. Result: Compared with the control group,Nao-shuning could significantly promote the proliferation of neural stem cells( P<0. 05)in a certain concentration range,which was dependent on the concentration,but the drug could inhibit stem cell proliferation and cause cell death in high concentrations. The results of WST-1 indicated that the Nao-shuning could proliferate the stem cells of MAPK/ERK signal pathway blocking treatment group,but the proliferation results were not significant. Western blot results showed that the Nao-shuning could significantly increase the expression of p-ERK1/2 in the MAPK/ERK signaling pathway and promote ERK1/2 phosphorylation. Conclusion: The above results suggested that the appropriate concentration of Chinese medicine Nao-shuning can promote the proliferation of neural stem cells cultured in vitro,and the mechanism may be related to promoting the phosphorylation of ERK1/2 in the MAPK/ERK signaling pathway.