Study On Active Ingredients And Therapeutic Mechanism Of Rheumatic Arthritis Of Artemisia Ordosica Based On Network Pharmacology Research Methods

Objectives:To systematically determine the chemical composition of the ethyl acetate and n-butanol extracts of Artemisia ordosica,to find out the overall chemical information of Artemisia ordosica by the combining the results of chemical composition of Artemisia ordosica petroleum ether and dichloromethane extracts with other researchers research and literature report chemical information analysis,to explore the mechanism of action of Artemisia ordosica by analyzing the overall chemical information of Artemisia ordosica and its network pharmacology research methods for the treatment of rheumatic arthritis.Methods:Conventional silica gel,Sephadex LH-20 and other column chromatography,TLC and high-performance liquid chromatography were used to separate and purify the ethyl acetate and n-butanol extracts of Artemisia ordosica to obtain a single compound.A single compound is determined and analyzed by various spectral data to determine its compound structural formula.At the same time,the structural formulas of all single compounds separated and refined and researched by the members of the research group were drawn using Chem draw 14.0 software and converted into standard Canonical SMILES format for storage.The SMILES format was imported into Swiss Target Prediction data to predict the target of the chemical composition of Artemisia ordosica.Related targets of rheumatic arthritis were searched in DisGeNET and OMIM databases.Integrating the intersection of the two targets,the STRING database is used to build a PPI network.The GO function enrichment and KEGG pathway enrichment analysis were conducted through DAVID,and then the Cytoscape 3.7.2 software was used to construct the compound-target-pathway network diagram.Results:15 compounds were isolated and identified from the ethyl acetate andn-butanol extracts of Artemisia ordosica,which were Integrin A(1),Integrin B(2),Ordosacid A(3),Ordosacid B(4),Ordosicin A(5),Ordosicin B(6),cinnamic acid(7),ferulic acide(8),caffeic acid(9),4,4'-dihydroxy-3,3'-dimethoxy-7-O-7'-epoxylignan(10),4,4',7,7'-tetrahydroxy-3,3'-dimethoxy-8-8'-lignan(11),4,4',7,7'-tetrahydroxy-3,3'-dimethoxy-7-O-7'-epoxylignan(12),Cymbadahoside A(13),Artemordolignan glycoside A'(14)and Physochloside A(15).According to the analysis of the overall chemical information of Artemisia ordosica,600 gene targets were obtained after excluding duplicates,and 55 targets were related to rheumatic arthritis.After enrichment analysis of GO function and KEGG path,281 BP entries,20 CC entries,36 MF entries and 35 KEGG signal paths were collected.After sorting by Degree and BC values,it was found that Ordosicin B,Integrin A,caffeic acid,ferulic acide,5-hydroxy-7,4'-dimethoxyflavone,etc.were the core components,and MAPK1,PIK3CG,PIK3CD,MMP9,MMP2,etc.were the core Target.Conclusion:12 compounds(1-12)were isolated and identified from the ethyl acetate extract of Artemisia ordosica,and 3 compounds(13-15)were isolated and identified from the n-butanol extract.Among them,compounds 1,2,3,4,5,6,14,15 are new compounds that have not been reported in the literature,and other compounds are isolated from this plant for the first time.The Mongolian medicine Artemisia ordosica can treat rheumatic arthritis through multiple components,multiple targets and multiple pathways.This study provides a new idea for the research on the mechanism of Artemisia ordosica treatment of rheumatic arthritis,and also points out the direction for the follow-up mechanism verification experiment.