目的探究丹龙醒脑方对脑缺血再灌注损伤大鼠海马区内源性神经干细胞增殖与β-catenin、Wnt-3a表达的影响。方法将60只雄性SD大鼠随机分为假手术组、模型组、依达拉奉组(西药组)、丹龙醒脑方小剂量组(丹小组)、丹龙醒脑方大剂量组(丹大组),线栓法制备局灶性脑缺血再灌注(I/R)损伤模型,再灌注7 d后取大鼠缺血侧脑组织。采用Brd U掺入法记录海马齿状回颗粒下区Brd U阳性细胞数目;采用RT-q PCR法检测海马总β-catenin、Wnt-3a m RNA表达;采用Western Blot法检测海马总β-catenin、胞浆β-catenin、Wnt-3a蛋白表达。结果与假手术组比较,其余各组Brd U阳性细胞数、β-catenin、Wnt-3a蛋白及m RNA的表达明显增强,差异有统计学意义(P〈0.05或P〈0.01);与模型组比较,西药组、丹小组、丹大组Brd U阳性细胞数、β-catenin、Wnt-3a蛋白及m RNA表达明显增强,差异有统计学意义(P〈0.05或P〈0.01)。结论丹龙醒脑方可能通过激活β-catenin及Wnt-3a表达促进神经干细胞增殖。
Objective To study the effect of Danlongxingnao recipe on proliferation of endogenous neural stem cells and expressions of β-catenin, Wnt-3a in hippocampus of cerebral ischemia-reperfusion injury model rats. Methods 60 male SD rats were randomly divided into sham operation group, model group, the edaravone group (western medicine group), Danlongxingnao recipe small dose group (Dan Xiao group), Danlongxingnao recipe high-dose group (Dan Da group). The focal cerebral ischemia reperfusion (I/R) injury models were prepared by suture method, and 7d after reperfusion took the brain tissue ischemia side of rats with reperfusion were taken out after 7d. BrdU incorporation method is used to record BrdU positive cells in the hippocampus dentate gyrus; Hippocampal β-catenin and Wnt-3a mRNA expressions were detected by RT-qPCR method; The hippocampus β-eatenin, cytoplasm β--catenin, Wnt-3a protein expression were determined by Western Blot method. Results Compared with sham operation group, the BrdU positive cells, 13-catenin,Wnt-3a protein and mRNA expression enhanced obviously in other groups, the difference was statistically significant (P〈0.05 or P〈0.01); Compared with model group, BrdU positive cells, β-catenin, Wnt-3a protein and mRNA expression improved obviously in western medicine group, Dan Xiao group, Dan Da group, the difference was statistically significant (P〈0.05 or P〈0.01). Conclusion Danlongxingnao recipe may promote the proliferation of neural stem cells by activating the Wnt-3a and β-catenin expressions