To Explore The Pathogenesis Of Purpuric Nephritis Based On Metabolomics And Network Pharmacology

Purpose:1.Apply LC-MS/MS metabolome analysis method to deeply study the pathogenesis and potential biomarkers of purpuric nephritis(HSPN).2.Use network pharmacology and molecular docking to explore the therapeutic mechanism of Qijishenkang Decoction(QJD)in the treatment of purpuric nephritis.Material and method:1.Metabonomics research materials and methods: From February 2019 to June 2019,21 children with purpuric nephritis aged 5-16 years were diagnosed and observed in the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine(receiving Qijishenkang Decoction,11 males and 10 female s).23 healthy children(11 males and 12females)were recruited through the Internet a nd on-site recruitment.The project passed the ethical review of clinical procedures(201 6(KT)-002-01),and completed clinical registration in clinical trials(registration number: NCT02878018).Obtained the informed consent form of the child guardian.Diagnosis basis: Diagnose HPSN according to the evidence-based medicine diagnosis and treatment guidelines for HSPN formulated by the Nephrology Branch of the Pediatric Branch of the Chinese Medical Association in2016,and the reference materials provided by Kidney Int Suppl.In this study,Qijishenkang Decoction was used to treat HSPN.Collect 2ml of morning urine from children with HSPN and healthy children and store it in a 5m L EP tube.Store it at-4°C for 30 min,then store it at-20°C for 24 h,an d finally move it to-80°C refrigerator In order to perform subsequent processing and analysis,avoid repeated freezing and thawing during storage,and use 150μL of 5% acetonitrile to reconstitute the sample before use,and wait for subsequent LC-MS/MS analysis.Before analyzing the actual sample,balance the system with a blank sample;during the analysis of the actual sample,every 10 stitches of the actual sample needs to be run 1 stitch of QC to monitor the pretreatment and the stability of the instrument operation.SIMCA(version 14.1,Umetrics AB,Umea Sweden)was used to perform pattern recognition analysis based on multivariate.2.Network pharmacology and molecular docking materials and methods:Relying on the Traditional Chinese Medicine Network Pharmacology Analysis Platform(TCMSP)to retrieve all the chemical components of the main drugs of Qijishenkang Decoction,use the Uniprot database to find the predicted and screened gene targets,and use the disease databases such as OMIM,GENECARDS and Dis Ge NET to construct the HSPN related target database,Using R language for network construction and GO and KEGG enrichment analysis,and at the same time,combined with metabolome research and molecular docking results to further verify the mechanism of Qijishenkang decoction in the treatment of HSPN.Results:1.LC-MS/MS metabolome results research shows that the pathogenesis of HSPN is cl osely related to human arginine biosynthesis,alanine,aspartic acid and glutamate meta bolism,and abnormal changes in the citric acid cycle.It is worth noting that this research group confirmed through clinical results and OPL S-DA chart that Qijishenkang Decoction can effectively treat children with HSPN,whi le the S chart results show that cystine and 1,3-dimethyluric acid are HSPN’s potential predictive biomarkers.2.The results of network pharmacology showed that: quercetin,luteolin,kaempferol,f ormononetin and isorhamnetin may be Qijishenkang decoction’s important ingredients that exert efficacy;PTGS2,PTGS1,F2,ESR1 and PRSS1 may be important targets of Qijishenkang decoction in the treatment of HSPN;GO and KE GG enrichment analysis show that the therapeutic effect of Qijishenkang decoction on HSPN may be similar to that of human body.The anti-inflammatory response,blood pressure regulation,arginine metabolism,platelet activation and Apelin signaling pathway are closely related;the successful docking of molecules demonstrates the possible mode of action of the active ingredients in QJD.Conclusion:1.The study of this experimental group found that the pathogenesis of HSPN may be related to abnormal changes in arginine biosynthesis.Cystine and 1,3-dimethyluric acid may be predictive biomarkers of HSPN.2.Combined analysis of the results of metabonomics and network pharmacology,this study found that the possible pathogenesis of HSPN and the therapeutic mechanism of Qijishenkang Decoction are focused on the human arginine metabolism pathway,for the follow-up in-depth study of the pathogenesis of HSPN provides a strong research f oundation.3.Through technical means such as network pharmacology and molecular docking,the metabolome research conclusions are further verified,which provides a reliable resear ch guidance for the in-depth study of Qijishenkang decoction on the treatment mechan ism of HSPN.The clinical benefit of treating children with HSPN provides an import ant reference basis.