| 1.BackgroundH.pyloriis a helicoid,microaerophilic gram-negative bacteria.It has been colonized in human gastric mucosa for a long time,and it’s the main pathogen of upper digestive tract.H.pyloriis known to be closely associated with the occurrence of gastrointestinal diseases such as chronic gastritis,gastric ulcer and gastric cancer,as well as extragastrointestinal diseases such as idiopathic purpura and iron deficiency anemia.Bismuth quadruple therapy,a classic therapy including bismuth,proton pump inhibitor and two kinds of antibiotics,is recommended by the guidelines for first-line treatment.However,the prevalence of bacterial antibiotic resistance and defective patient compliance appear to be evolving over time,and subsequently,the eradication rate of H.pyloriis decreasing.Also,the gstric microbiota was disrupted by antibiotics and cause several adverse events like diarrhea,constipation and abdominal pain.Further studies into the development of new treatment based on the bismuth quadruple therapy are required.Nowadays,probiotics have been used in the clinical practice,making it to become the hotspot and trend of H.pyloriinfection treatment and prevention.2.Methods and ResultsMethods:(1)The transcriptomics used in gene-set enrichment analysis(GSEA)and weighted gene co-expression network analysis(WGCNA)were downloaded from gene expression omnibus(GEO)database.GSEA and WGCNA were used to find the hub modules and pathways related to H.pyloriinfection.(2)The mRNA level of enriched genes from bioinformatic analysis was verified by real time-qPCR in cells.Proteomics was performed on human gastric mucosa tissues to explore the change of genes and pathways in protein level and the molecular mechanism of H.pyloriinfection.(3)The high throughput 16S rRNA sequencing was performed to detect the alteration in microbiota on gastric mucosa from patients with H.pyloriinfection or not.(4)WGCNA was used to explore the probiotics which had negative correlation with H.pylori,and Oxford Cup method was applied to verify the probiotics with the capability of inhibit H.pylori.Results:(1)There were 3 modules and 25 KEGG pathways enriched in the analysis of GSEA and WGCNA,including B cell receptor signaling pathway and cell adhesion molecule signal pathway,etc.(2)The results of real time-qPCR from GES-1 cells with H.pyloriSSl stain infection showed that 80 genes were up regulated,21 genes were down regulated,and the rest 15 genes remain unchanged.The results of proteomics showed that 71 differentially expressed proteins were isolated,and the they could be enriched into several signal pathways,such as the cell adhesion molecules signal pathway.(3)The results of 16S rRNA sequencing showed that the H.pylorifamily was the dominant family in microbiota,the diversity and richness of the gastric microbiota were decreased.The LEfSe analysis showed that the Lactobacilli,Streptococcaceae and Bacteroideae in the H.pyloriinfection group were lower than those in the control group and they might be the potential probiotic strains for the treatment of H.pyloriinfection.(4)The results of WGCNA and LEfSe analysis showed that Lactobacillaceae and other 10 families were down regulated and negatively correlated with H.pylori.The Oxford Cup method proved that Lactobacillus rhamnosus LRX-01 inhibits H.pylori.3.Conclusion(1)3 modules and 25 pathways related with H.pyloriinfection were screened through GSEA and WGCNA anlysis.(2)Real time-qPCR and proteomics were used to verify the results of bioinformatic analysis at transcription and protein levels.An amount of 80 genes,3 KEGG pathways were verified and 71 differentially expressed proteins were identifed.(3)High throughput sequencing indicated that H.pyloriinfection increases the proportion of H.pyloriin the stomach mucosa and decrease the diversity of bacterial flora.(4)Bioinformatc analysis predicted that 11 families of microbiota have the potential to inhibit H.pyloriand Lactobacillus rhamnosus LRX-01 had an antimicrobial efficacy against H.pylori. |
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