基于UPLC-Q-TOF-MS/MS和网络药理学研究连翘治疗急性肺损伤的作用机制

Study on the mechanism of Fructus Forsythiae in the treatment of acute lung injury based on UPLC-Q-TOFMS/MS and network pharmacology

目的:基于UPLC-Q-TOF-MS/MS技术分析连翘的入血成分,并结合网络药理学研究连翘治疗急性肺损伤(acute lung injury,ALI)的作用机制。方法:使用脂多糖(lipopolysaccharide,LPS)建立ALI大鼠模型,采用UPLC-Q-TOF-MS/MS技术鉴定连翘的入血成分,结合PharmMapper、GeneCards和Disgenet数据库筛选连翘入血成分治疗ALI的潜在作用靶点,String平台绘制蛋白互作(PPI)网络图,Cytoscape软件做网络拓扑分析并筛选核心靶点,通过David数据库对核心靶点富集分析,使用Autodock Vina和PyMol软件对相关入血成分与核心靶点进行分子对接。结果:肺组织切片结果显示ALI大鼠模型制备成功;得到37个连翘入血成分;连翘治疗ALI核心靶点70个,涉及PI3K/Akt、MAPK和Ras等152个相关通路;agatholic acid、无梗五加苷B、连翘苷、连翘酯苷A和芦丁5个入血成分与核心靶点JAK2和JAK3分子对接的结合能均小于-4.25 kJ·mol-1,对接结果良好。结论:该研究明确了连翘的入血成分,网络药理学预测agatholic acid、无梗五加苷B、连翘苷、连翘酯苷A和芦丁可能为连翘防治ALI的主要成分,其作用机制可能与靶向JAK2和JAK3调控PI3K/Akt信号通路有关。

OBJECTIVE To study the mechanism of Fructus Forsythiae in the treatment of acute lung injury(ALI)based on UPLC-Q-TOF-MS/MS technology and network pharmacology.METHODS The rat model of ALI was established by lipopolysaccharide(LPS).The entering blood components of Fructus Forsythiae were identified by UPLC-Q-TOF-MS/MS technology.The potential targets of Fructus Forsythiae in the treatment of ALI were screened by combining PharmMapper,GeneCards and Disgenet.The protein interaction(PPI)network diagram was drawn on String platform.The network topology was analyzed by Cytoscape,and the core targets were screened.The core targets were enriched and analyzed by David.Autodock Vina and PyMol were used for molecular docking between the relevant blood components and the core targets.RESULTS The results of lung tissue section showed that the ALI rat model was successfully obtained;thirty-seven entering blood components of Fructus Forsythiae were obtained;Fructus Forsythiae had seventy core targets for ALI,involving one hundred and fifty-two related pathways such as PI3K/Akt,MAPK and RAS;the binding energies of agatholic acid,acanthoside B,phillyrin,forsythoside A and rutin to the core targets JAK2 and JAK3 were less than-4.25 kJ·mol^(-1),and the docking results were good.CONCLUSION In this study,the entering blood components of Fructus Forsythiae were identified.Network pharmacology predicted that agatholic acid,acanthoside B,phillyrin,forsythoside A and rutin might be the main components in the prevention and treatment of ALI with Fructus Forsythiae,and its mechanism might be related to the regulation of PI3K/Akt signaling pathway by targeting JAK2 and JAK3.