心脑宁胶囊通过调节NF-κB介导的抑炎通路保护脑缺血再灌注损伤的作用及机制研究

Protective effect and mechanism of Xinnaoning Capsule on cerebral ischemia-reperfusion injury by regulating NF-κB mediated anti-inflammatory pathway

关键词:  脑缺血再灌注  心脑宁胶囊  核转录因子-κB

Key words:  cerebral ischemia reperfusion  Xinnaoning Capsule  NF-κB

摘要:

[目的] 研究心脑宁胶囊对脑缺血再灌注损伤的保护作用。[方法] 雄性ICR小鼠随机分为假手术组、模型组、心脑宁胶囊组和依达拉奉组。给药7 d后，制备小鼠中动脉闭塞模型，缺血30 min，再灌注24 h。进行神经功能损伤、组织水肿、梗死面积和组织病理损伤的评价；利用网络药理学分析心脑宁胶囊作用于脑缺血再灌注损伤的核心靶点；采用蛋白免疫印迹法和免疫组化法测定核转录因子-κB（NF-κB）p65蛋白的表达。[结果] 心脑宁胶囊组神经行为学评分、中线偏移距离、梗死面积均显著低于模型组（P<0.01）；苏木精-伊红染色结果显示，心脑宁胶囊组较模型组病理损伤有所改善；网络药理学分析显示，心脑宁胶囊作用机制与RelA（p65）密切相关；蛋白免疫印迹结果显示，心脑宁胶囊组NF-κB p65总蛋白的表达较模型组明显降低（P<0.01）；免疫组化染色结果显示，心脑宁胶囊组阳性细胞数较模型组明显减少（P<0.01），采用积分光密度测量法，与模型组之间的差异更为显著（P<0.001）。[结论] 心脑宁胶囊在大脑中动脉阻塞模型小鼠中通过下调NF-κB的表达及活化降低炎症反应，改善脑缺血再灌注损伤。

Abstract:

[Objective] To observe the effect of Xinnaoning Capsule on cerebral ischemia reperfusion injury.[Methods] Male ICR mice were randomly divided into sham-operation group,model group,Xinnaoning group and Edaravone group. After the pre-dosed 7 days,a mouse middle artery occlusion (MCAO) model was established,and ischemia for 30 minutes,reperfusion for 24 h. The evaluation of nerve function damage,tissue edema,infarct area and histopathological damage were carried out in sequence;the core target of Xinnaoning Capsule's active components on cerebral ischemia reperfusion injury was analyzed by network pharmacology;Western blot and immunohistochemistry were used to determine the expression of nuclear transcription factor-B (NF-κB p65).[Results] The neurobehavioral score,brain midline dislocation distance,and infarct area of the Xinnaoning group were significantly lower than those of the model group (P<0.01);the results of hematoxylin-eosin (HE) staining showed that compared with the model group,the Xinnaoning group was improved in terms of pathological damage;network pharmacological analysis shows that the mechanism of Xinnaoning capsules is closely related to RelA (p65);Western blot results show that the expression of total NF-κB p65 protein in Xinnaoning group was significantly lower than that of model group (P<0.01);the results of immunohistochemical staining showed that the number of positive cells in the Xinnaoning group was significantly less than that of the model group (P<0.01). the difference with the model group was more significant if the integral optical density measurement method was used,(P<0.001).[Conclusion] Xinnaoning Capsule effectively improved cerebral ischemia-reperfusion injury in MCAO mice via at least in part by an anti-inflammatory mechanism indicated by reduction of the expression and activation of NF-κB.