摘要
目的探讨王不留行黄酮苷对小鼠脑缺血性脑损伤的影响。方法通过GeneCard、Swiss Target Prediction、Targetnet、Pharmmapper和TTD等数据库分别对王不留行黄酮苷和缺血性脑损伤的靶点进行预测并取二者交集(检索日期:2024年10月30日);通过Sting平台建立交集靶点的蛋白相互作用(PPI)网络;采用Cytoscape软件进行PPI网络可视化处理;通过DAVID数据库和微生信在线基因功能富集分析工具对王不留行黄酮苷和缺血性脑损伤的交集靶点进行基因本体论(GO)和全基因组及代谢途径(KEGG)富集分析;利用AutodockVina软件将王不留行黄酮苷与PPI网络图中核心靶点蛋白进行分子对接。在动物实验中,通过制备小鼠大脑中动脉远端闭塞模型模拟脑缺血,造模后给予腹腔注射10/20/40 mg/kg的王不留行黄酮苷5 d,并分别在造模后1,3,5 d进行神经功能评分和转棒实验,第5天给药结束后进行2,3,5-氯化三苯基四氮唑(TTC)染色和病理切片染色,验证王不留行黄酮苷对小鼠脑缺血损伤的作用。结果通过各数据库检索去重取交集后获得王不留行黄酮苷和缺血性脑卒中(IS)的交集靶点,利用Sting平台建立交集靶点的PPI网络共获取63个交集靶点;GO富集和KEGG富集分析结果显示,王不留行黄酮苷主要对缺氧反应、炎症和凋亡等生物过程进行调控,并且可能通过血管内皮生长因子(VEGF)和白介素(IL)-17等信号通路作用于缺血性脑卒中;分子对接结果显示其与degree值≥40的核心靶点都有很强的结合能,其结合能均<-7 kcal/mol。动物实验表明,王不留行黄酮苷可以降低脑缺血小鼠的改良神经功能缺损(mNSS)评分,延长其在棒时间,差异均具有统计学意义(P<0.05或P<0.01);与假手术组相比,王不留行黄酮苷干预后可明显减小小鼠脑梗死体积,差异具有统计学意义(P<0.05或P<0.01);病理切片染色也表明,王不留行黄酮苷可明显抑制神经元数量减少并改善神经元细胞的形态;尼氏小体数量丢失也明显减少。结论王不留行黄酮苷能够有效减轻小鼠缺血后脑组织损伤,其可能通过多通路多靶点对缺血性脑损伤发挥保护作用。
Objective To investigate the neuroprotective effect of vccarin on ischemic brain injury in mice.Methods Potential targets of vaccarin and ischemic brain injury were predicted using databases such as GeneCard,Swiss Target Prediction,Targetnet,Pharmmapper,and TTD(search date:October 30,2024).Intersecting targets were identified and analyzed via protein-protein interaction(PPI)network construction(STRING database;visualization with Cytoscape).Functional enrichment analysis(GO and KEGG pathways)was performed using DAVID and MicrobeDB.Molecular docking(AutoDock Vina)assessed binding affinities between vaccarin and core targets(degree≥40).In animal experiments,focal cerebral ischemia was induced in mice via distal middle cerebral artery occlusion(dMCAO).Vaccarin(10,20,40 mg/kg)or vehicle was administered intraperitoneally for 5 days post-surgery.Neurological function scores and rotarod tests were performed on days 1,3,and 5 post-surgery.After drug administration on day 5,triphenyltetrazolium chloride(TTC)staining and histopathological section staining were performed to verify the effects of vaccarin on ischemic brain injury in mice.Results After removing duplicates and obtaining the intersection of the targets,a total of 63 overlapping targets of vaccarin and ischemic stroke(IS)were identified.The PPI network of the intersecting targets was constructed using the Sting platform.GO and KEGG enrichment analysis indicated that vaccarin primarily regulated biological processes such as hypoxia response,inflammation,and apoptosis,and Highlighted VEGF and IL-17 signaling pathways.Molecular docking results showed that the binding energies of vaccarin with core targets(degree≥40)were strong,with binding energies all less than-7 kcal/mol.In animal experiments,vaccarin significantly reduced the mNSS neurological scores and prolonged the time spent on the rotarod in ischemic mice,with statistically significant differences(P<0.05 or P<0.01).Compared to the sham group,vaccarin intervention significantly reduced the infarct volume in mice,with statistical significance(P<0.05 or P<0.01).Pathological staining showed that vaccarin significantly alleviated the loss of neurons and improved neuronal morphology.The loss of Nissl bodies was also notably reduced.Conclusion Vaccarin confers neuroprotection in ischemic brain injury by modulating hypoxia-inflammatory-apoptosis pathways via multi-target interactions,with VEGF and IL-17 signaling as key mechanistic components.
作者
孙攀喜
魏丽丽
陈勇
樊东升
Sun Panxi;Wei Lili;Chen Yong;Fan Dongsheng(Department of Neurology,Peking University Third Hospital,Beijing 100191,China;Shihezi University School of Medicine,Xinjiang Key Laboratory of Regional and Ethnic High-incidence Diseases,Ministry of Education,Shihezi 832000,China)
出处
《中华脑血管病杂志(电子版)》
2025年第3期220-227,共8页
Chinese Journal of Cerebrovascular Diseases(Electronic Edition)
基金
国家自然科学基金项目(81471184)
北京大学第三医院院重点项目(BYSYZD2021004)。
