基于PI3K/Akt/NOS通路探讨鞣花酸在脑缺血再灌注损伤大鼠的神经保护作用

Neuroprotective effect of ellagic acid on cerebral ischemia-reperfusion injury in rats based on PI3K/Akt/NOS pathway

目的基于磷酯酰激醇3-激酶/蛋白激酶B/一氧化氮合酶(PI3K/Akt/NOS)通路探讨鞣花酸对脑缺血再灌注损伤大鼠的神经保护作用。方法将36只雄性Wistar大鼠按数字表法随机分为3组假手术(Sham)组、缺血再灌注组和鞣花酸预干预组,线栓法制作大鼠右侧大脑中动脉缺血120 min再灌注模型。鞣花酸预干预为造模前14 d按100 mg/(kg·d)给予鞣花酸灌胃,每组12只。运用神经功能缺损评分法评估各组动物的行为学变化,HE染色观察各组大鼠缺血侧脑组织病理改变,Western blotting法检测缺血侧脑组织凋亡蛋白及PI3K/Akt/NOS信号通路蛋白。结果(1)鞣花酸可以提高缺血再灌注后神经功能评分(P<0.05)。(2)模型组大鼠缺血侧脑组织凋亡相关蛋白(Bax/Bcl-2、CytC、Cleaved caspase-3)水平比Sham组明显升高(P<0.05),鞣花酸组大鼠缺血侧脑组织凋亡相关蛋白(Bax/Bcl-2、CytC、Cleaved caspase-3)水平比模型组组明显减少(P<0.05)。(3)模型组大鼠缺血侧脑组织中(p-Akt/Akt及p-eNOS/eNOS)比Sham组明显降低(P<0.05),鞣花酸组大鼠缺血侧脑组织中(p-Akt/Akt及p-eNOS/eNOS)明显升高(P<0.05)。结论鞣花酸可以激活PI3K/Akt通路减轻大鼠脑缺血再灌注损伤,保护脑组织。

Objective To investigate the neuroprotective effect of ellagic acid on rats with cerebral ischemia-reperfusion injury based on the pathway of phosphatidylinositol 3-kinase/protein kinase B/nitric oxide synthetase(PI3K/Akt/NOS).Methods 36 male Wistar rats were randomly divided into three groupssham group,ischemia-reperfusion group,and ellagic acid pre-intervention group.The right middle cerebral artery ischemia-reperfusion model was made by thread bolt method.In the pre-intervention of ellagic acid,12 rats in each group were given ellagic acid by gavage 14 days before modeling.The behavioral changes of the three groups were evaluated by neurological deficit score,The pathological changes of the ischemic brain were observed by HE staining.The apoptotic protein and PI3K/Akt/NOS signal pathway protein were detected by Western blotting.Results Ellagic acid can improve the neurological score after ischemia-reperfusion(P<0.05).The level of Bax/Bcl-2,cytc and cleaved caspase-3 in the model group was significantly higher than that in the sham group(P<0.05).The level of Bax/Bcl-2,cytc and cleared caspase-3 in the ellagic acid group was significantly lower than that in the model group(P<0.05).In model group,p-Akt/Akt and p-eNOS/eNOS were significantly lower than those in sham group(P<0.05),while in ellagic acid group,p-Akt/Akt and p-eNOS/eNOS were significantly higher(P<0.05).Conclusion Ellagic acid can activate PI3K/Akt pathway to alleviate cerebral ischemia-reperfusion injury and protect brain tissue.