益气活血方和补肾生髓方对局灶性脑缺血再灌注大鼠海马Wnt3a、Frizzled9和TCF3表达的影响

Effects of Yiqi Huoxue formula and Bushen Shengsui formula on expression of Wnt3a,Frizzled9 and TCF3 in hippocampus of rats with ischemia-reperfusion injury

目的探讨益气活血方和补肾生髓方对脑缺血大鼠海马Wnt/β-catenin信号通路Wnt3a、卷曲蛋白9(Frizzled9)和T细胞因子3(TCF3)蛋白表达的影响。方法采用线栓法制作局灶性脑缺血大鼠模型,随机分为假手术组、模型组、益气活血方组和补肾生髓方组,脑缺血2 h后持续灌注7d。采用Western-Blot法检测缺血侧海马Wnt3a、Frizzled9和TCF3蛋白表达,用免疫组化Envision两步法检测缺血侧海马CA1区Wnt3a、Frizzled9和TCF3蛋白表达。结果在缺血侧海马区,与假手术组比较,模型组Wnt3a、Frizzled9和TCF3蛋白的相对表达量均显著增加(P<0.01);与模型组比较,益气活血方组Wnt3a、Frizzled9和TCF3蛋白的相对表达量均显著降低(P<0.01),补肾生髓方组Wnt3a、Frizzled9和TCF3蛋白的相对表达量均显著降低(P<0.01或P<0.05)。在缺血侧海马CA1区,与假手术组比较,模型组Wnt3a、Frizzled9和TCF3蛋白表达显著增加(P<0.01);与模型组比较,益气活血方组与补肾生髓方组能显著降低CA1区Wnt3a、Frizzled9和TCF3蛋白表达(P<0.01)。结论 2种中药复方能通过抑制缺血侧海马Wnt3a、Frizzled9和TCF3蛋白表达,调节Wnt/β-catenin信号通路,促进局灶性脑缺血再灌注损伤脑组织修复。

Objective To study the effects of Yiqi Huoxue formula （YQHX） and Bushen Shengsui formula （BSSS） on the protein expression of Wnt3a,Frizzled 9 and TCF3 on Wnt/β-catenin pathway in hippocampus of rat model induced by middle cerebral artery ischemia/reperfusion. Methods The focal cerebral ischemia/reperfusion rat model was established by intraluminal thread occlusion of the middle cerebral artery （MCAO）for 2-h followed by 7-d reperfusion. The rats were randomly divided into sham group, model group, YQHX group and BSSS group（each n = 10）. The expression of Wnt3a, Frizzled9 and TCF3 protein in hippocampus of cerebral ischemia were detected by Western-Blot assay, and those in CA1 zone of cerebral ischemia were measured by using immunohistochemistry Envision two-step method. Results The relative expression of Wnt3a, Frizzled 9 and TCF3 protein in both hippocamous and CAl zone increased significantly in model group compared with sham group （ P 〈0.01 ）. Compared with model group, the expression of those three proteins in both hippocamous and CA1 zone decreased significantly in YQHX group and BSSS group （ P 〈 0.05 ）. Conclusion Yiqi Huoxue formula and Bushen Shengsui formula can promote the repairment of injured cerebral tissue by the way of reducing the protein expression of Wnt3a, Frizzled9 and TCF3 in hippocampus to regulate Wnt/β-eatenin signal pathway.