丹龙醒脑方对脑缺血再灌注大鼠侧脑室室管膜下区神经干细胞增殖及Hes1、Hes5表达的影响

Effects of Danlong Xingnao Formula on Proliferation of Neural Stem Cells in Sub Ventricular Zone and Expressions of Hes1 and Hes5 in Cerebral Ischemia-Reperfusion Injury Model Rats

目的观察丹龙醒脑方对脑缺血再灌注模型大鼠侧脑室室管膜下区(SVZ)神经干细胞(NSCs)增殖与Hes1、Hes5表达的影响,探讨其促进内源性NSCs增殖的作用机制。方法将80只雄性SD大鼠随机分为假手术组、模型组、依达拉奉组(依达组)、丹龙醒脑方组(丹龙组)。采用线栓法制备局灶性脑缺血再灌注模型,再灌注7 d后取缺血侧SVZ脑组织。Brdu免疫荧光法检测SVZ NSCs增殖,RT-q PCR、Western blot分别检测Hes1、Hes5 m RNA和蛋白的表达。结果与假手术组比较,其余各组Brdu阳性细胞率增加,Hes1、Hes5 m RNA及蛋白表达明显升高(P<0.01);与模型组比较,依达组、丹龙组Brdu阳性细胞率明显增加,Hes1、Hes5 m RNA及蛋白表达水平明显增强(P<0.01);丹龙组Hes1 m RNA表达水平优于依达组(P<0.01),其余指标均无明显差异。结论丹龙醒脑方可促进脑缺血再灌注后大鼠SVZ NSCs增殖,并上调Hes1、Hes5表达水平,其机制可能与激活Notch信号通路有关。

Objective To study effects ofDanlong Xingnao Formula (DLXNF) on proliferation of neural stem cells (NSCs) and the expressions of Hes1 and Hes5 in sub ventricular zone (SVZ) in cerebral ischemia-reperfusion injury model rats; To explore the mechanism of promoting the proliferation of NSCsMethods Eighty male SD rats were randomly divided into sham-operation group, model group, edaravone group andDLXNF group. The focal cerebral ischemia reperfusion injury models were prepared by suture method, and 7 d after reperfusion, the SVZ brain tissue of ischemia side was taken. The proliferation of cells was detected by Brdu labeling fluorescence immunocytochemistry; Hes1, Hes5 mRNA and protein expressions were detected by fluorescence real-time quantitative PCR and Western blot method in each group.Results Compared with the sham-operation group, Brdu positive cell rate in other groups increased more obviously, and the expressions of Hes1, Hes5 mRNA and protein also increased significantly (P<0.01). Compared with the model group, Brdu positive cell rate increased significantly in edaravone group and DLXNF group, and the expressions of Hes1, Hes5 mRNA and protein increased significantly (P<0.01). The expression of Hes1 mRNA in DLXNF group was superior to that in edaravone group (P<0.01), and other indexes had no significant difference.Conclusion DLXNF can promote the proliferation of NSCs in SVZ in cerebral ischemia-reperfusion injury model rats, and up-regulate the expressions of Hes1 and Hes5, whose mechanism may be related to the activation of Notch signaling pathway.