Transcriptomics And Network Pharmacology Of SAPHO Syndrome

Objective:To identify differentially eepressed genes related to SAPHO syndrome and explore the "multi-targeted" curative mechanism of Tripterygium wilfoirdii for SAPHO syndrome,so as to pro,vide mechanistic evidence for clinical application of Tripterygium wilfordii preparations in the treatment of SAPHO syndrome.Materials and methods:(1)Six patients with SAPHO syndrome and six healthy volunteers were recruited for the identification of differentially expressed genes in patients with SAPHO syndrome based on whole transcriptome sequencing of peripheral blood neutrophils.Co-expression analysis and target-predictive analysis of non-coding RNAs were performed and network of competitive endogenous RNA was constructed.DAVID tool website was used for function annotation of differentially expressed genes based on KEGG signaling pathway and GO term enrichment analysis.The differentially expressed genes were verified by qRT-PCR with an enlarged sample size,and correlated with clinical indicators to detect the predictive ability of differentially expressed genes for SAPHO syndrome.(2)Main components(OB>30%,DL>0.18)and corresponding targets of Tripterygium wilfordii were screened through TCMSP database,symmap database and literature review.A protein-protein interaction(PPI)network between component targets and SAPHO-related genes was then constructed based on STRING database.Enrichment of KEGG signaling pathway and GO Terms of genes in the PPI network were carried out using DAVID tool website.And finally,Triptolide,one of the main active ingredients of Tripterygium wilfordii,was co-cultured with neutrophils purified from patients with SAPHO syndrome to verify its effect on predicted signal pathways and biological processes.Flow cytometry,indirect immunofluorescence as well as Trans well assay were conducted to evaluate the effect of Triptolide on neutrophil apoptosis,NETosis and cell migration.Result:Whole transcriptome sequencing identified 442 differentially expressed genes in SAPHO syndrome,of which 294 were up-regulated and 148 down-regulated.Functional enrichment analysis indicated that differentially expressed genes were related to cell adhe sion and cell migration.A total of 2713 differentially expressed long-chain non-coding RNAs were identified,including 1030 up-regulated genes and 1683 down-regulated genes.Based on Pearson correlation coefficient,a co-expression network of lncRNA-RNA was constructed including 281 nodes,of which 139 were differentially expressed mRNAs and 142 were differentially expressed.lncRNAs Small RNA sequencing identified 136 small non-coding RNAs,of which 36 were up-regulated and 100 were down-regulated.The target mRNAs of SAPHO syndrome-related miRNAs were predicted,and 40491 genes were obtained.These genes can be rich in signal pathways in KEGG database,including adhesion plaques,vascular endothelial growth factor,insulin resistance and cancer-related signaling pathways.Fifty-eight main components(OB>30%,DL>0.18)and 198 corresponding targets of Tripterygium wilfordii were collected according to TCMSP database,symmap database and literature review.A protein-protein interaction(PPI)network between targets of Tripterygium wilfordii components and SAPHO-related genes was then constructed based on STRING database.Enrichment of KEGG signaling pathways and GO terms of nodes in the PPI network indicated that components of Tripterygium wilfordii may benefit patients with SAPHO syndrome by influencing neutrophil apoptosis,NETosis and migration,etc.One of the main ingredients of Tripterygium wilfordii,Triptolide,was selected for the verification of predicted function.In vitro,Triptolide enhanced apoptosis but inhibited NETosis and cell migration of activated neutrophils from patients with SAPHO syndrome.Conclusion:In conclusion,this study provides a first insight into PPI network between targets of Tripterygium wilfordii components and SAPHO-related genes,and we suppose that Tripterygium wilfordii may function by interfering with cell apoptosis,NETosis and migration of neutrophils from patients with SAPHO syndrome.