黄芪多糖缓解急性脑缺血再灌注损伤的氧化应激反应和免疫功能紊乱

Astragalus polysaccharide alleviates oxidative stress response and immune dysfunction of acute cerebral ischemia reperfusion injury

目的:探索黄芪多糖(AP)对急性脑缺血再灌注损伤的氧化应激反应和免疫功能紊乱的影响。方法:采用大脑中动脉线栓法建立缺血再灌注损伤模型。苏木素伊红(HE)染色分析组织病理学变化。Tunel染色检测病变组织细胞凋亡。试剂盒检测组织中氧化应激相关指标的含量;ELISA检测血清中炎症因子的含量。蛋白印迹检测B淋巴细胞瘤-2(Bcl-2)、BAX、炎性复合体隐热蛋白(NLRP3)、凋亡相关斑点样蛋白(ASC)、cl-caspase-1的表达。结果:模型组与对照组相比,脑缺血再灌注大鼠脑组织出现严重病变,细胞凋亡明显增加,同时Bcl-2表达显著下调,BAX表达显著上调;组织中SOD的活力明显下降,GSH的浓度显著降低,MDA的浓度显著上升;血液中IL-6、TNF-α、IL-1β、IL-18的含量显著上升;脑组织中NLRP3、ASC、cl-caspase-1的表达显著下调(P<0. 01)。加药组与模型组相比较,急性脑缺血再灌注大鼠脑组织病变减弱,细胞凋亡明显减少,Bcl-2蛋白水平显著上调,BAX蛋白水平显著下调;组织中SOD的活力和GSH的浓度显著上升,MDA的浓度显著下降;血液中IL-6、TNF-α、IL-1β、IL-18的含量显著降低;脑组织中NLRP3、ASC、cl-caspase-1的表达显著下调(P<0. 01);且随着黄芪多糖加药浓度的增加,效果越明显。结论:黄芪多糖通过影响细胞自噬和免疫相关蛋白的水平,缓解急性脑缺血再灌注损伤的氧化应激反应和免疫功能紊乱。

Objective:To explore the effects of astragalus polysaccharide on oxidative stress response and immune dysfunction in acute cerebral ischemia reperfusion injury.Methods:The model of ischemia reperfusion injury was established by middle cerebral artery occlusion.The histopathological changes were analyzed by hematoxylin-eosin(HE).Tunel staining was used to detect the apoptosis of diseased tissue cells.The oxidative stress indicator content were detected by related kit.ELISA was used to detect the concentration of inflammatory factor.The expression level of Bcl-2,BAX,NLRP3,ASC,cl-caspase-3 were detected by Western blot.Results:Compared with the control group,cerebral ischemia reperfusion rats had serious pathological changes.The number of apoptosis positive cells was increased significantly,while the expression of Bcl-2 was significantly down-regulated,the expression of BAX was significantly up-regulated.The activity of SOD in the tissue decreased significantly,the concentration of GSH decreased significantly,and the concentration of MDA increased significantly.In the blood,IL-6,TNF-α,IL-1βand IL-18 increased significantly.The expression of NLRP3,ASC and clcaspase-1 in brain tissue was significantly reduced(P<0.01).Compared with the model group,the cerebral tissue lesion in rats with acute cerebral ischemia was decreased,the apoptosis was significantly reduced,and the level of Bcl-2 protein was significantly increased,and the expression level of BAX protein was significantly down-regulated.The activity of SOD in the tissue and the concentration of GSH increased significantly,and the concentration of MDA decreased significantly.In the blood,IL-6,TNF-α,IL-1βand IL-18 were significantly reduced.The expression of NLRP3,ASC and cl-caspase-1 in brain tissues was significantly reduced(P<0.01).And with the increase of astragalus polysaccharide concentration,the more obvious effect.Conclusion:Astragalus polysaccharide alleviates oxidative stress response and immune dysfunction of acute cerebral ischemia reperfusion injury by affecting the level of autophagy and immune-re-lated proteins.