黄芪-灯盏细辛组分最佳配比对脑缺血大鼠的保护作用与机制

Protective effect and mechanism of Astragalus combined with Erigeron breviscapus in the best proportion on cerebral ischemia rats

目的:确证黄芪-灯盏细辛注射液联用最佳配比的治疗效果,探讨药物治疗脑缺血的作用机制。方法:采用线栓法制作大鼠大脑中动脉缺血再灌注模型,将大鼠分为黄芪注射液单用组,灯盏细辛注射液单用组,黄芪∶灯盏细辛=4∶6组,模型组和假手术组,大鼠脑缺血24h和48h后进行神经行为学评分,测定其脑梗死率、血清丙二醛(MDA)含量、超氧化物歧化酶(SOD)活力,并对缺血48h大鼠大脑皮层进行免疫组化测定AKT、P-AKT蛋白表达和Western blot检测AKT、Bad蛋白表达。结果:与模型组相比,24h和48h后给药各组均有治疗脑缺血的效果,神经行为学评分明显降低,脑梗死率明显减少,MDA含量均明显减少,SOD活力均明显升高,48h后各组免疫组化测定AKT和P-AKT均明显升高,WB检测中给药组AKT上调,Bad下调,其中黄芪∶灯盏细辛=4∶6组(即黄芪注射液0.8ml/kg,灯盏细辛注射液1.2ml/kg)的治疗效果显著。结论:黄芪注射液、灯盏注射液对脑缺血大鼠有治疗效果,黄芪-灯盏注射液联用对大鼠脑缺血的治疗作用效果最佳,其机制可能是刺激PI3K/AKT信号通路升高AKT、P-AKT信号蛋白表达和抑制Bad蛋白表达有关。

Objective:To study the therapeutic effect of Astragalus-Erigerontis Injection with the best prescription,and to explore its mechanism on cerebral ischemia.Methods:Ischemia-reperfusion of middle cerebral artery rat model was established by suture method.All rats were divided into astragalus injection group,Erigeron Injection group,astragalus:Erigeron=4:6 combination injection group,the model group and the sham operation group.After 24 and 48 hours of the surgery,rats in each group were scored by neurobehavioral assessment.After 48 hours,the cerebral infarction rate,serum malondialdehyde(MDA)content and superoxide dismutase(SOD)activity were determined.The expression levels of AKT,P-AKT protein were determined by immunohistochemistry,the protein expression levels of AKT and Bad were determined by western blot.Results:Compared with the model group,drugs showed therapeutic effect on cerebral ischemia 24 h and 48 h after surgery.The neurobehavioral score,cerebral infarction rate and the content of MDA were decreased,the level of SOD was increased.After 48 hours,protein levels of AKT and P-AKT were increased by immunohistochemistry In western blot assay,AKT level was increased and Bad was significantly inhibited.Astragalus:Erigeron=4:6 combination injection showed the best effect(astragalus 0.8 ml/kg and Erigeron1.2 ml/kg).Conclusion:Both Astragalus membranaceus and Erigeron breviscapus have therapeutic effects on rat cerebral ischemia,and the combination injection has the best effect.The mechanism may be related to stimulating PI3 K/AKT signal pathway to increase the expressions of AKT and P-AKT signaling proteins and inhibit Bad protein expression.