| Objective: To construct the ’ target-pathway ’ network of Zhuang medicine Qingdu Shenjin Decoction(QDSJF)regulating rheumatoid arthritis through network pharmacology,and to verify the core target by combining with collagen-induced arthritis(CIA)rat experiment,to explore the mechanism of QDSJF and provide clinical application basis.Methods: 1.The main active compounds were screened by TCMSP database with drug-likeness ≥ 0.18 and oral bioavailability ≥ 30% as the conditions,and the target names were standardized by using the certified human genes in Uniprot database,and the related targets were converted into corresponding drug targets.2.Input ’ rheumatoid arthritis ’ content,collect all targets on rheumatoid arthritis in the Gene Cards database and OMIM database as a disease data set.3.The ’ disease-compound-component-gene target ’ regulatory network was constructed by Cytoscape,and the main active components of QDSJF were identified by the evaluation parameters calculated by network topology.Then the NCC algorithm in the plug-in Cyto Hubba was used to calculate the core targets in all intersection targets.4.The metascape database was used for gene ontology function and Kyoto Encyclopedia of Genes and Genomes pathway annotation.5.The binding energy was calculated by Autodock Tools1.5.6 molecular docking software to evaluate the affinity of the main active ingredients and the core target.6.The rat model of collagen-induced arthritis(CIA)was constructed as the research object,and the prediction results of network pharmacology were preliminarily verified by rat animal model.In this study,70 SD female rats were randomly divided into normal group,model group,negative control group,QDSJF high dose group,medium and low dose group and methotrexate group,10 rats in each group.During the experiment,the hair,food intake,water intake,body weight and activity of all rats were observed,and the levels of key proteins in peripheral blood of rats were detected by enzyme-linked immunosorbent assay.Results: 1.From the analysis of network pharmacology,145 active components and 238 drug targets in qingdushenjin decoction were obtained,and 151 purpose were combinatied with rheumatoid arthritis.2.The components were β-sitosterol,pedicelflavin,stigmasterol,luteolin and quercetin.MMP9,VEGFA,JUN,TNF,IL6,CASP3,STAT3,AKT1,PTGS2 and IL1β are the fatal targets.3.In consequences of molecular docking demonstrate that the leading purpose components and vital targets had nice combination.4.From the analysis set up by the experimental group,there were significant differences(P<0.01)between the model group and the negative control group and the normal group at the levels of MMP9,AKT1 and PTGS2,indicating that the experimental detection indicators were comparable;and the negative control group had no significant difference in the above protein levels compared with the model group(P>0.05),indicating that the influence of gavage administration on the detection indicators of this experiment could be excluded.5.In consequence,elisa assay testify that there was a statistically sense differently within the administration group,the model group,the negative control group and the normal group(P<0.01),explanatory note that the establish model was prosperous.There were significant differences in MMP9 and AKT1 between each dose group of qingdushenjin decoction and methotrexate group and negative control group and model group(P<0.01).There was no prominent property difference in PTGS2 between the low-dose group and the model group and the negative control group(P > 0.05).Conclusion: 1.Network pharmacology and molecular docking predict that Qingdushenjin prescription for RA may act on STAT3,AKT1,PTGS2,MMP9,VEGFA,JUN,TNF,IL6,CASP3,IL1 β and other core targets in TN through main active compounds such as mastermasterol,β-sitosterol,quercetin,luteolin and formononetin F signaling pathway and IL17 signaling pathway play multi-component,multi-target and multi-pathway roles.By constructing the top 3 targets of molecular docking binding ability in CIA rats,the results showed that Qingdu Shenjin formula could improve the symptoms of affected joints in CIA rats and significantly reduce the levels of MMP9,AKT1 and PTGS2.The main mechanism may be firstly to reduce the proliferation of synovial cells by inhibiting PTGS2,and improve the aggregation of inflammation under hypoxia conditions in joints.Secondly,it can reduce the MMP9 level in the joints and protect the articular cartilage.Then,it acts on AKT1-targeted chondrocyte apoptosis to control the progression of ossification,and exerts the therapeutic mechanism together.2.QDSJF is based on the theory of "poison deficiency causes all diseases" in Zhuang medicine,and has definite and unique curative effect in the treatment of rheumatoid arthritis,which can provide medication reference for the diagnosis and treatment of this disease. |
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