Research On The Pharmacodynamic Material Basis And Mechanism Of Anti-Rheumatoid Arthritis From Tradition Chinese Medicine Based On Virtual Screening

Rheumatoid Arthritis(RA)is a chronic disease with unknown etiology,which seriously affects the life of patients.Although the commonly used drugs can improve the condition,they need to be used for a long time and have various side effects.Therefore,it is urgent to develop new anti-rheumatoid arthritis drugs.As a unique medical resource in China,traditional Chinese medicine has a long history in the treatment of RA diseases,which provides ideas for the discovery of new drugs against RA.In this study,the virtual screening technology was used to screen a series of compounds on the related targets of RA,such as cyclooxygenase-2(COX-2)and C-JUN amino terminal kinase(JNK)from the natural product database,and found some drug molecules with good activity,meanwhile,the material basis and mechanism of action against rheumatoid arthritis were further explored,which provided a theoretical basis for the discovery of new drugs in the treatment of rheumatoid arthritis.The specific research contents and conclusions are as follows:1.Screening target proteins based on pharmacophore model: The pharmacophore model belonged to the cyclooxygenase-2(COX-2)and c-jun amino terminal kinase(JNK)signal receptor protein target of rheumatoid arthritis was constructed.It was found that the related target proteins had important pharmacodynamic characteristic elements.2.Research on virtual screening based on molecular docking: the binding energy values of active molecules and inactive molecules were calculated by Autodock vina program,so as to draw ROC curve and compound histogram,calculate EF value and hit rate of active molecules.The results showed that the molecular docking models of 3ln1 and 4awi target proteins had strong enrichment ability of active molecules.Using the molecular docking model of the above target proteins,large-scale virtual screening was carried out on the natural product database,and a total of three traditional Chinese medicine components with binding energy lower than the threshold,molecular structure similar to the original ligand and meeting the five principles of drug-like were obtained.3.Lymphocyte proliferation experiment: the mouse spleen lymphocytes were selected as the experiment object,and three active Chinese medicine molecules obtained were used to carry out proliferation of spleen lymphocytes in mice by CCK-8 experiment.It is found that sennoside D and kaempferol had obvious effects on lymphocyte proliferation.Two traditional Chinese medicine ingredients(sennoside D and kaempferol)played a important role in anti-RA through reducing proliferating lymphocytes.4.Reverse molecular docking verification and network pharmacology research: 35 potential action targets of traditional Chinese medicine active components against rheumatoid arthritis were obtained through mapping the targets of two traditional Chinese medicine components with disease targets.The action mechanism of above two active components against rheumatoid arthritis was also explored based on network pharmacology.The results showed that the two active components mainly acted on the key targets of JUN and TNF,and regulated the TNF,PI3K-Akt,HIF-1,MAPK,T cell receptor,NOD and other signaling pathways.to play a role in anti-rheumatoid arthritis.In addition,the molecular docking results showed that the two active components of traditional Chinese medicine had good binding affinity with the two key targets,which further explained the reliability of network pharmacology prediction and molecular docking results.To sum up,this paper used the target proteins screened by pharmacophore model to conduct virtual screening based on molecular docking,and found a number of traditional Chinese medicine components with potential anti rheumatoid arthritis.The leading compounds with obvious anti-rheumatoid arthritis effect were obtained through cell proliferation experiment,and the action mechanism of the active components of traditional Chinese medicine against rheumatoid arthritis was explored by the research strategies of network pharmacology and reverse molecular docking,which provided a new idea and direction for discovering the active components of anti-rheumatoid arthritis from traditional Chinese medicine.