Study On The Identification Of Material Basis And Anti-rheumatoid Arthritis Mechanism Of Chaenomelis Fructus

Objectives: To clarify the main chemical components of Chaenomelis Fructus,predict its main pharmacodynamic targets through network pharmacology and verify the pharmacodynamic targets with metabolomics and molecular biology so that the mechanism of Chaenomelis Fructus' s pharmacodynamic effects could be clarified.Methods: 1.The main chemical constituents in Chaenomelis Fructus were analyzed by building a database according to existing literature,using ultra-high liquid chromatography-quadrupole-electrostatic orbitrap mass spectrometry technology,library search combined with standard identification and mass spectrometry identification.On this basis,the prototype components of blood were analyzed.2.Incorporated the compounds identified in Chaenomelis Fructus and the chemical components of blood into the analysis,conducted a component-target interaction network study,predicted the component targets by searching the database,and screened the potential targets of Chaenomelis Fructus treatment for rheumatoid arthritis.The topology analysis,protein-protein interaction(PPI)network analysis and GO and KEGG enrichment analysis were used to analyze the component-target network to predict the target metabolic pathways.The study on the chemical component-target interaction network of Chaenomelis Fructus into the blood provided a direction for further study of the Chaenomelis Fructus' s efficacy and mechanism of action.3.In response to the inflammatory metabolic pathways and core targets predicted by network pharmacology,the rats' right feet were injected with Freund 's complete adjuvant(FCA)to induce arthritis,and the toe volume of the rats was measured by using toe volume measuring instrument.The weight of mice was measured by using a weight scale.The levels of IL-1?,IL-6,and TNF-? in the serum before and after Chaenomelis Fructus intervention and the expression of RA model rats joint synovium Bax,Bcl-2,Fas,ERK,JNK,P-ERK,P-JNK protein.At the same time,its efficacy and mechanism were verified through metabolomics technology.Results: 1.By using ultra-high liquid chromatography-quadrupole-electrostatic orbital trap mass spectrometry technology,the structures of 47 chemical components were identified from Chaenomelis Fructus,mainly including triterpenoids,flavonoids and organic acids.Nine prototype ingredients were found in the components of blood,including 7 organic acids and 2 flavonoids,indicating that organic acids were the main prototype ingredients of the components of blood in Chaenomelis Fructus.2.The component-disease-target network study of 47 chemical components screened out 42 targets for Chaenomelis Fructus treatment of rheumatoid diseases.The intersection of blood components-targets and diseases-targets was taken to screen out 27 targets in Chaenomelis Fructus for treatment of rheumatoid diseases.The core targets are PTGS2,MMP9,SRC,TNF,IL6,IL1 B and so on.Enrichment analysis of core targets GO and KEGG showed that they were mainly involved in IL-17 signaling pathway,arachidonic acid metabolism and positive regulation of inflammation.3.After different doses of Chaenomelis Fructus alcohol extract intervening in RA model rats,the swelling of the rats' planta decreased to varying degrees in the treatment groups of the low,medium and high dose of Chaenomelis Fructus extract,and the arthritis score gradually were improved.The effect of the medium and high dose treatment group was the most significant,showing a good anti-inflammatory effect.HE staining results showed that the inflammation changes in each Chaenomelis Fructus alcohol extract group were improved in different degrees compared with the model group in terms of inflammatory cell infiltration and synovial cell hyperplasia.ELISA test results showed that Chaenomelis Fructus alcohol extract could significantly reduce the levels of inflammatory factors IL-1?,IL-6 and TNF-? in rat serum and increase the level of IL-10.The RA model rats' joints synovial Bax,Bcl-2,Fas,ERK,JNK,P-ERK,and P-JNK protein expression disorder was significantly improved after the intervention of Chaenomelis Fructus alcohol extract.Metabolomics studies found that Chaenomelis Fructus mainly intervened in arachidonic acid metabolism and branched-chain amino acid catabolism,thereby inhibiting the production of inflammatory factors and reducing the release of inflammatory mediators in order to achieve anti-inflammatory effects.Conclusion: With significant anti-inflammatory effect,the main component of Chaenomelis Fructus' s anti-inflammatory is organic acids.Its anti-inflammatory effect is mainly performed through the intervening in serum inflammatory factors IL-1?,IL-6,TNF-? levels,increasing IL-10 level and the expression of Bax,Bcl-2,Fas,ERK,JNK,P-ERK,P-JNK protein in the joint synovium and the intervening in the arachidonic acid metabolism and branched-chain amino acid catabolism.