Assessing The Anti-inflammatory Mechanism Of Reduning Injection Based On Systems Pharmacology Methods

ObjectiveThis study is expected to establish a real-time dose-response model to describe the regulatory characteristics of Reduning injection on the inflammatory process through systems pharmacology methods like network analysis,mathematical modeling and experimental verification.By exploring the pharmacokinetic properties of the main pharmacodynamic components of Reduning injection,the real-time dose-response relationship of Reduning injection in regulating the inflammatory process was described,so as to reveal its anti-inflammatory mechanism at the systematic level and provide a new method for the mechanism research of traditional Chinese medicine.Method1 By consulting references and data mining,the main pharmacodynamic compounds and metabolites of Reduning injection were collected.Relevant information and threedimensional structure files were collected from PubChem.Targets of the main pharmacodynamic compounds and their metabolites of Reduning injection were collected through database retrieval(PubChem,BindingDB,DrugBank and TCMSP)and molecular docking(autodock 4.2.6).A compound-target network(CTN)was constructed and visualized using Cytoscape,and the topological parameters of the network were calculated to screen out the key targets and main active compounds of Reduning Injection in the treatment of inflammation.Relevant pathways of targets of Reduning Injection were collected from KEGG.Through the construction and analysis of target-pathway network(TPN)and literature mining,key signaling pathways of Reduning Injection was determined.2 An HPLC method for the quantitative analysis of main pharmacodynamic compounds of Reduning Injection in rat plasma was establish.The plasma concentrations of these compounds were measured at different time points after the rats were injected with high,medium and low dose Reduning injection via tail vein.The pharmacokinetic parameters of each compound were calculated by non atrioventricular model.The pharmaceutical-time curves was drawn and the pharmacokinetic behavior of each compound was studied.3 Molecular docking was used to predict the targets of main active components of Reduning injection.We collected the signaling pathways related to the targets from KEGG,explored the relationship between these pathways and inflammation,and established a crosstalk network.Through further analysis and evolution,the mechanism network of Reduning Injection regulating inflammation was obtained.Network efficiency and network flux were used to evaluate the effect of Reduning Injection on the mechanism network.Enzyme-linked immunosorbent assay(ELISA)was used to detect the relative reduction of inflammatory factors in LPS induced inflammatory model rats after receiving Reduning injection,and the evaluation model was verified.Results1 Data mining and molecular docking were used and 11 active components,84 targets and 201 signaling pathways were screened out to be related to the anti-inflammatory activity of Reduning injection.Through network analysis,it was found that 4 key targets(Bcl-2,eNOS,PTGS2 and PPARα)and 4 key pathways(estrogen signaling pathway,PI3K-Akt signaling pathway,cGMP PKG signaling pathway and calcium signaling pathway)would play important roles in the therapeutic effect of Reduning injection in treating inflammatory diseases.2 An HPLC method for the simultaneous determination of 10 main active components of Reduning Injection in SD rat plasma was established for the first time.The methodological investigation of this method meets the requirements,and it has high sensitivity,good selectivity,good precision and reproducibility.The method has been successfully applied to the pharmacokinetic studies of the main active components in Reduning injection.3 Molecular docking was used to screen out a total of 13 targets of 10 main active compounds of Reduning injection.Combined with database mining and references,a regulatory mechanism network of inflammation was established,which was composed of 137 nodes and 297 edges.The real-time dose-response relationship of Reduning Injection on the regulation of inflammation after intravenous injection was quantitatively evaluated by using network efficiency and network flux and the real-time changes of blood concentration of each compound.The reliability of the model was verified by ELISA.ConclusionIn this work,the "multi-component multi-target multi-pathway”effect of Reduning injection on inflammatory proccess was explored by systems pharmacology.An inflammatory mechanism network was constructed.An HPLC method was established for the simultaneous determination of 10 main active components in Reduning injection.The pharmacokinetics of main active components in Reduning injection were studied.A network model was used to simulate the expression of various inflammatory factors after administration,and ELISA was used to verify the simulation results.In this study,the antiinflammatory mechanism of Reduning injection was explored by systems pharmacology,which provides a new method for the study of anti-inflammatory mechanism of Reduning injection as well as the mechanism of traditional Chinese medicine.