益气活血化浊解毒方对脑缺血再灌注损伤大鼠脑组织VEGF、Notch1、Dll4表达的影响

Effects of Yiqi Huoxue Huazhuo Jiedu recipe on the expression of VEGF, Notch1 and Dll4 in brain tissue of rats with cerebral ischemia reperfusion injury

目的观察益气活血化浊解毒方对脑缺血再灌注损伤大鼠的治疗作用,初步探讨其保护神经血管单元的机制。方法将SPF级大鼠随机分为假手术组、模型组、中药低剂量组、中药中剂量组、中药高剂量组和尼莫地平组。采用线栓法制备大脑中动脉闭塞模型,各组予相应干预,3天后计算各组大鼠神经功能评分,TTC染色测定脑梗死面积,免疫组化检测大鼠脑组织血管内皮生长因子(vascular endothelial growth inhibitor,VEGF)蛋白表达,q-PCR检测大鼠脑组织Notch1、Dll4 mRNA表达,Western blot检测大鼠脑组织VEGF、Notch1、Dll4蛋白表达。结果(1)神经功能评分结果显示:中药低、中、高剂量组及尼莫地平组较模型组大鼠神经功能缺失评分均有所降低(P<0.05);(2)脑梗死面积结果显示:中药低、中、高剂量组及尼莫地平组较模型组大鼠脑梗死面积显著缩小(P<0.05);(3)免疫组化结果显示:中药低、中、高剂量组及尼莫地平组较模型组大鼠脑组织VEGF蛋白表达显著增大(P<0.05);(4)q-PCR结果显示:中药低、中、高剂量组及尼莫地平组较模型组大鼠脑组织Notch1、Dll4 mRNA表达明显增加(P<0.05)(5)免疫蛋白印迹法结果显示:中药低、中、高剂量组及尼莫地平组较模型组大鼠脑组织VEGF、Notch1、Dll4蛋白表达增加(P<0.05)。结论益气活血化浊解毒方可以调控脑缺血再灌注损伤大鼠神经血管单元发挥神经保护作用,其机制可能与上调大鼠脑组织VEGF、Notch1、Dll4表达有关。

Objective To observe the therapeutic effect of Yiqi Huoxue Huazhuo Jiedu recipe on cerebral ischemic reperfusion injury in rats,and to explore its mechanism of protecting neurovascular unit.Methods SPF rats were randomly divided into sham operation group,model group,Chinese medicine low-dose group,Chinese medicine medium-dose group,Chinese medicine high-dose group and nimodipine group.Three days later,the neurological function score was observed,the cerebral infarction area was measured by TTC staining,the expression of VEGF protein in rat brain tissue was detected by immunohistochemistry,the expression of Notch1 and Dll4 mRNA in rat brain tissue was detected by q-PCR,and the protein expressions of VEGF,Notch1 and Dll4 in brain tissue were detected by Western blot.Results(1)The results of neurological deficit scores shows that the Chinese medicine low,middle,and high dose groups and the nimodipine group had significant reduced in neurological deficit scores(P<0.05).(2)The results of cerebral infarction area showed that the Chinese medicine low,middle and high dose group and nimodipine group were significantly smaller than the model group(P<0.05).(3)The results of immunohistochemistry showed that compared with the model group,the expression of VEGF protein in the Chinese medicine low,medium,and high dose groups and the Nimodipine group was significantly increased(P<0.05).(4)The results of q-PCR showed that the expressions of Notch1 and Dll4 mRNA in the Chinese medicine low,medium and high-dose groups and nimodipine group were significantly higher than the model group(P<0.05).(5)The results of Western blot showed that compared with the model group,the expressions of VEGF,Notch1,and Dll4 protein in brain tissue of the Chinese medicine low,medium,and high dose groups and the Nimodipine group were significantly increased(P<0.05).Conclusion Yiqi Huoxue Huazhuo Jiedu recipe can regulate the neurovascular units of rats with cerebral ischemia reperfusion injury and play a neuroprotective role.The mechanism may be related to the up-regulation of VEGF,Notch1 and Dll4 expression in the brain tissue of rats.