基于UPLC-Q-TOF-MSE的青紫合剂成分分析及其治疗过敏性紫癜的作用机制预测和实验验证

Component analysis of Qingzi Mixture based on UPLC-Q-TOF-MSE, and prediction and experimental verification of its mechanism in treating Henoch-Sch?nlein Purpura

关键词:  青紫合剂  过敏性紫癜  UPLC-Q-TOF-MSE  网络药理学  分子对接

Key words:  Qingzi mixture  Henoch-Sch?nlein purpura  UPLC-Q-TOF-MSE  network pharmacology  molecular docking

摘要:

目的 分析青紫合剂的化学成分，探讨其治疗过敏性紫癜(Henoch-Schonlein purpura，HSP)的网络药理学作用机制并验证。方法 利用UPLC-Q-TOF-MSE技术结合UNIFI软件鉴定青紫合剂的化学成分。运用Seaware、SwissTargetPrediction数据库预测鉴定所得成分的作用靶点；同时在GeneCards、Disgenet等数据库获取HSP的作用靶点。利用STRING数据库分析靶点蛋白质-蛋白质相互作用关系，利用Metascape数据库进行GO功能富集分析和KEGG通路富集分析。利用SYBYL-X 1.2软件对活性成分与作用靶点进行分子对接。采用蛋白质印迹法检测青紫合剂对热证结合卵蛋白诱导的HSP模型大鼠关键通路的影响。结果 在青紫合剂中共鉴定出150种化学成分。成分靶点与疾病靶点交集62个。GO功能富集和KEGG通路分析显示，PI3K/Akt、MAPK、HIF1A信号通路为关键通路，STAT3、EGFR、CXCL8为关键靶点；分子对接结果显示5个活性成分与核心靶点的结合活性较好。动物实验发现，青紫合剂可显著改善HSP大鼠的皮肤损伤，降低血清异常IgA1及炎症因子水平，抑制皮肤PI3K/Akt通路的蛋白表达。结论 青紫合剂中的槲皮素、阿魏酸、高香草酸、异阿魏酸、原儿茶酸等可能为其治疗HSP的主要成分，可能是通过调控PI3K/Akt信号通路发挥作用。

Abstract:

OBJECTIVE To identify the chemical composition of Qingzi mixture, and to explore and verify its mechanism in treating Henoch Schonlein purpura (HSP) through network pharmacology. METHODS UPLC-Q-TOF-MSE technology was combined with UNIFI analysis software to quickly identify the components in Qingzi mixture. Seaware and SwissTargetPrediction database were used to predict the targets of the identified compounds. GeneCards and Disgenet database were used to screen the disease targets of HSP. Target protein-protein interactions were analyzed using the STRING database, GO functional enrichment analysis and KEGG pathway enrichment analysis were performed using the Metascape database. Molecular docking analysis was conducted by SYBYL-X 1.2 software to verify the binding activity between active ingredients and targets. Western blot was used to detect the effect of Qingzi mixture on key pathways in HSP model rats induced by heat syndrome combined with ovalbumin. RESULTS A total of 150 compounds were identified in Qingzi mixture. 62 intersection of component targets and disease targets were screened. The results of GO function annotation and KEGG signaling pathway analysis showed that PI3K/Akt, MAPK, HIF1A pathway were the key pathways, among which STAT3, EGFR, CXCL8 were the key targets. Molecular docking showed that the five active ingredients of Qingzi mixture had robust binding activities with the core targets. Animal experiments showed that Qingzi mixture significantly improved skin damage in HSP rats, reduced the serum levels of abnormal IgA1 and inflammatory factors, and inhibited the protein expression of the PI3K/Akt pathway in skin tissues. CONCLUSION Quercetin, ferulic acid, homovanillic acid, isoferulic acid, protocatechuic acid of Qingzi mixture might be the main substances of Qingzi mixture for treating HSP, which may play a role in regulating PI3K/Akt signaling pathway.