Study On Mechanism Of Treating Henoch-sch?nlein Purpura With Xiao'er Zidian Zhenxiao Granules

Objective: To explore the mechanism of treating Henoch-Sch?nlein Purpura(HSP)with Xiao'er Zidian Zhenxiao granules.Methods: 1.Analysis of the Mechanism of in Treating Allergic Purpura Based on Network Pharmacology:Based on TCMSP and Batman-TCM,the chemical constituents and targets of Xiao'er Zidian Zhenxiao granules were retrieved.The targets of Henoch-Schonlein Purpura were collected by Genecard,and then the targets of Xiao'er Zidian Zhenxiao granules were obtained by integrating data.Cytoscape 3.2.1 was used to construct the network of Xiao'er Zidian Zhenxiao granules-Active Compound-Therapeutic Target.String was used to construct the PPI network of Xiao'er Zidian Zhenxiao granules,and the mechanism of Xiao'er Zidian Zhenxiao granules in the treatment of Henoch-Schonlein Purpura was studied.2.Analysis of Xiao'er Zidian Zhenxiao Granules by Using UPLC-Q-Orbitrap-MS :Agilent SB C18(3.0mm×100 mm,1.8 ?m)column was used with 0.1% formic acid solution(A)-acid acetonitrile(B)as the mobile phase for gradient elution.The high-resolution quadrupole orbitrap mass spectrometer was used as detector with electrospray ion source.3.The metabolomics research method based on LC-MS technology was used to analyze the urine of children with HSP and healthy children.The effects of pediatric allergic purpura on the changes of urine metabolites in children were analyzed by UPLC-Q-TOF / MS analysis Biomarkers of Allergic Purpura.4.(1)A retrospective analysis of the relevant test indicators of 171 children with HSP was conducted to investigate the general condition,month of onset,morbidity,concomitant disorders,general inflammation indicators,infection of Mycoplasma pneumoniae and chlamydia,immunoglobulin and complement C3 levels,Respiratory virus infection and other related indicators.(2)Collect early morning fasting serum from children with HSP and use Enzyme Linked ImmunoSorbent Assay(ELISA)to detect the IL-6,IL-8,TNF-?,IgA1,IgA / Fc?RI(CD89)values before and after treatment.(3)HUVECs cells were cultured with F12 k medium containing serum from children with HSP and intervened with Pediatric Purpura Xiaoxiao Granules.ELISA method was used to detect IL-6,IL-8,TNF-?,IgA / Fc?RI(CD89)values,Westernblot Detection of Syk,p-Syk,PI3 K,p-PI3 K protein expression.Results: 1.The network diagram of Xiao'er Zidian Zhenxiao granules-active compound-therapeutic target included 9 single drugs,206 active compounds and 40 therapeutic targets.PPI network contains 39 target proteins,and the key proteins involve IL6,INS,and VEGFA.There are 14 entries of gene ontology(GO),which involve the reaction of lipopolysaccharide,hypoxia,drug and so on.The Kyoto Encyclopedia of Genes and Genomes(KEGG)contains four pathways,including tumor necrosis factor signaling pathway,NF-kappa B signaling pathway,HIF-1 signaling pathway and PI3K-AKT signaling pathway.2.Based on the analysis of mass spectrometry and literature reports,38 compounds were confirmed,including 10 flavones,4 anthraquinones,2 terpenes,2 shikonofurans,1steroline,1 polyphenol and 1 lignan.3.The metabolites in 92 urine samples were selected as biomarkers of allergic purpura disease in children,and 15 of them were identified,which may be the difference between children with allergic purpura and healthy children.4.(1)The general inflammation index of children with HSP in the acute phase has increased,suggesting that infections such as bacteria,viruses,and Mycoplasma pneumoniae may be the cause of the development of allergic purpura.The levels of serum immunoglobulins IgM,IgG,IgA,and complement C3 in children with HSP are present Disorders,the serum IgA and complement C3 levels were significantly higher in the HSP group than in the normal control group;respiratory virus infections in children with HSP were mainly adenovirus,human respiratory syncytial virus,human parainfluenza virus,and coxsackie virus.(2)The serum inflammatory cytokines IL-6,IL-8,and TNF-? in children with acuteHSP were all reduced after treatment compared with before treatment.The decrease in IgA1,IgA / Fc?RI(CD89)was statistically significant(P <0.05).(3)The serum levels of IL-6,IL-8,TNF-?,IgA / Fc?RI(CD89)in HUVECs supernatants increased after serum stimulation in children with HSP(P <0.05),and their expression levels decreased after intervention with Xiao'er Zidian Zhenxiao granules(P<0.05).Conclusion: 1.The results of this study preliminarily predicted the pharmacological action and molecular mechanism of Xiao'er Zidian Zhenxiao granules,and laid a theoretical foundation for further study of the pharmacological action and molecular mechanism of Xiao'er Zidian Zhenxiao granules in the treatment of Henoch-Schonlein purpura.2.The UPLC-Q-Orbitrap-MS method is simple,reliable and rapid to identify the chemical compositions of Xiao'er Zidian Zhenxiao Granules,and it is helpful to reveal its chemical constituents and pharmacodynamic substances.3.The difference of metabolic markers in urine between children with allergic purpura and healthy children may provide a method for disease identification,syndrome type diagnosis and disease treatment.4.The role of Xiao'er Zidian Zhenxiao granules in the treatment of allergic purpura may be related to the targeted regulation of IgA/Fc purpura RI against allergic purpura vascular endothelial cell inflammation.