基于网络药理学探讨Win55,212-2对GP120诱导大鼠艾滋性认知功能障碍的影响及其机制

Study on the effect and mechanism of Win55,212-2 on GP120-induced HAND in rats based on network pharmacology

目的:基于网络药理学探讨Win55,212-2对艾滋病病毒-1(HIV-1)包膜蛋白GP120诱导大鼠艾滋性认知功能障碍(HAND)的影响及其机制。方法:通过GeneCards、OMIM、DisGeNET和NDCI数据库获取HAND相关靶点,GeneCards、Pharm Mapper、SwissTargetPrediction数据库获取Win55,212-2相关靶点,再通过Venny 2.1.0在线工具映射获得HAND与Win55,212-2的交集靶点。采用String 11.5数据库和Cytoscape 3.7.2软件构建交集靶点的蛋白相互作用(PPI)网络并筛选出核心靶点;通过David 6.8数据库对交集靶点进行GO和KEGG通路富集分析,并通过微生物信息学在线工具对富集结果进行可视化。最后采用Cytoscape 3.7.2软件构建“药物—靶点—疾病—通路”网络。通过脑部立体定位注射技术建立GP120诱导的HAND大鼠模型,术后第3天进行新物体识别测试,检测各组大鼠的认知功能。实时荧光定量PCR(RT-qPCR)检测各组大鼠海马组织中P38 MAPK通路和下游炎症因子TNF-α、CXCL-12 mRNA表达水平。结果:Win55,212-2与HAND的共同作用靶点81个,GO和KEGG分析结果显示Win55,212-2治疗HAND的核心靶点包括AKT1、TNF、ALB、MAPK3等;主要通路涉及cAMP信号通路、AGE-RAGE信号通路、TNF信号通路、MAPK信号通路等。新物体识别测试结果显示,Win55,212-2提高了GP120诱导HAND大鼠的辨别指数,RT-qPCR结果显示Win55,212-2降低了P38、TNF-α、CXCL-12的mRNA表达水平(均P<0.01)。结论:Win55,212-2通过下调P38 MAPK通路并抑制下游炎症因子TNF-α、CXCL-12的表达而起到神经保护作用,这可能是其改善GP120诱导大鼠HAND的机制。

Objective:To explore the effect and mechanism of Win55,212-2 on human immunodeficiency virus type 1(HIV-1)envelope protein GP120-induced HIV-1-associated neurocognitive disorders(HAND)in rats based on network pharmacology.Methods:First,HAND related targets were obtained from GeneCards,OMIM,DisGeNET and NDCI databases,and Win55,212-2 related targets were obtained from GeneCards,Pharm Mapper and SwissTargetPrediction.Then,the intersection targets of HAND and Win55,212-2 were obtained by using Venny 2.1.0 online tool.The protein interaction(PPI)network of intersection targets was constructed by String 11.5 database and Cytoscape 3.7.2 software,and the core targets were screened out.GO and KEGG pathway enrichment analysis was performed for intersection targets using David 6.8 database,and the enrichment results were visualized using bioinformatics softwares.Finally,a network model diagram of"drug-target-disease-pathway"was constructed by using Cytascape 3.7.2 software.The rat model of GP120-induced HAND was established by stereotactic injection in-to the brain,and the new object recognition test(NORT)was performed on the third day after surgery to detect the cognitive function of each group of rats.Real-time fluorescence quantitative polymerase chain reaction(RTqPCR)was used to detect the mRNA expression levels of P38 MAPK pathway and downstream inflammatory factors TNF-a and CXCL-12 in the hippocampus of rats in each group.Results:There were 81 common targets of Win55,212-2-HAND,and the results of GO and KEGG analysis showed that the core targets of Win55,212-2 for HAND treatment included AKT1,TNF,ALB and MAPK3,etc.The main pathways involved cAMP signaling pathway,AGE-RAGE signaling pathway,TNF signaling pathway and MAPK signaling pathway,etc.The new object recognition test(NORT)results showed that Win55,212-2 increased the discrimination index of GP120-induced HAND rats,and RT-qPCR results showed that Win55,212-2 decreased the mRNA expression levels of P38,TNF-αand CXCL-12(all P<0.01).Conclusion:Win55,212-2 plays a neuroprotective role by down-regulating P38 MAPK pathway and inhibiting the expression of downstream inflammatory factors TNF-αand CXCL-12,which may be the mechanism of improving GP120-induced HAND in rats.