基于网络药理学和动物实验探讨白藜芦醇抗脊髓损伤作用机制

To Explore the Mechanism of Resveratrol Against Spinal Cord Injury Based on Network Pharmacology and Animal Experiments

目的:使用网络药理学方法和动物实验探讨白藜芦醇抗脊髓损伤的作用机制。方法:使用TCMSP搜集及Swiss平台预测,获得白藜芦醇作用靶点。使用Gene card、OMIM、TTD、Drug bank数据库搜集脊髓损伤相关靶点。韦恩图对二者取交集,即为白藜芦醇抗脊髓损伤的作用靶点。利用STRING网站和cytoscape软件构建交集靶点蛋白的互作网络图(PPI)。根据网络中拓扑参数的大小筛选关键作用靶点。使用DAVID网站进行GO和KEGG分析。通过免疫组化、RT-qPCR实验验证白藜芦醇对脊髓损伤模型大鼠中部分关键靶点的调控作用。结果:共获得白藜芦醇抗脊髓损伤作用靶点48个,其中关键作用靶点8个即AKT1、TNF、VEGFA、IL-1B、IL-6、JUN、CASP3、HIF1A。KEGG分析富集显著的通路主要有:TNF信号通路、PI3K-Akt信号通路、IL-17信号通路等。动物实验发现,白藜芦醇对脊髓损伤后大鼠运动功能具有明显改善作用,并可明显干预AKT1、TNF、VEGFA mRNA表达。结论:白藜芦醇可能通过作用于AKT1、TNF、VEGFA等靶点,调控TNF信号通路、PI3K-Akt信号通路、IL-17信号通路等发挥抗脊髓损伤作用。

Objective:To explore the mechanism of resveratrol against spinal cord injury by using network pharmacological methods and animal experiments.Methods:TCMSP collection and Swiss platform were used to obtain the action target of resveratrol.Use Gene card,OMIM,TTD,and Drug bank databases to collect targets related to spinal cord injury.The intersection of the two is the action target of resveratrol against spinal cord injury.The interaction network diagram(PPI)of the intersection target proteins was constructed by using STRING website and cytoscape software.According to the size of the topological parameters in the network,the key targets are selected.Use DAVID website for GO and KEGG analysis.Immunohistochemical and RT-qPCR experiments were used to verify the regulatory effect of resveratrol on some key targets in rats with spinal cord injury.Results:A total of 48 anti-spinal cord injury targets of resveratrol were obtained,including 8 key targets,namely AKT1,TNF,VEGFA,IL-1B,IL-6,JUN,CASP3,HIF1A.The pathways with significant enrichment in KEGG analysis mainly include TNF signal pathway,PI3K-Akt signal pathway,IL-17 signal pathway,etc.Animal experiments showed that resveratrol could significantly improve the motor function of rats after spinal cord injury,and could interfere with the expression of AKT1,TNF,VEGFA mRNA.Conclusion:Resveratrol may act on AKT1,TNF,VEGFA and other targets,regulate TNF signal pathway,PI3K-Akt signal pathway,IL-17 signal pathway and play an anti-spinal cord injury role.