Mechanism Of IL-17 In Regulating Gastric Epithelial LCN2-induced Helicobacter Pylori Infection

Objective:To investigate whether Helicobacter pylori infection affects LCN2 expression and whether its upregulation is regulated by IL-17 cytokines.Method:Using bioinformatics to verify the rationality of LCN2 gene selection,the keyword "Helicobacter pylori infection" was searched in the GEO database,and 1271 results were obtained,and 3 suitable datasets were screened: GSE27411,GSE10262,GSE74492,GEO2 R was used to analyze the expression of LCN2 genes in the difference between infected and non-infected group samples in three datasets.By plotting the Wayne plot,the differential genes that were jointly upregulated by the three datasets were identified,and the relationship between LCN2 was explored.Subculture of the Sydney standard strain SS1 for subsequent animal model establishment and cell experiments for infection;A mouse model of Helicobacter pylori infection was established,verified in vivo by mouse experiments,a healthy control group and a Helicobacter pylori infection group were set up,and the expression of IL-17 in the suspension of mouse spleen cells was measured by flow cytometry between the two groups.The gastric mucosa of the two groups was immunohistochemical and HE stained to observe the expression and inflammation of LCN2 in the gastric mucosa of the two groups.The anti-IL-17 control group was added to measure the expression of LCN2 in the blood of the three groups of mice by ELISA technology,and the differential expression of LCN2 in the three groups was observed.In vitro verification was carried out by cell experiments,GES-1 cells were co-cultured with Helicobacter pylori,and IL-17 antibodies and their blocking antibodies were added to the co-culture system to observe whether the secretion of LCN2 would rise or decrease under antibody interference.Outcome:The bioinformatics results showed that the expression of LCN2 in the samples infected with Helicobacter pylori was significantly higher than that in the non-Helicobacter pylori infection group,which verified the rationality of LCN2 gene selection.After subculturing,the Sydney standard strain was verified and collected smoothly in large quantities;In mouse experiments,The secretion of IL-17 cytokines in mice after Helicobacter pylori infection increased significantly(P<0.05),not only the degree of inflammation was also significantly increased;the results of ELISA and immunohistochemistry showed that the degree of LCN2 infiltration in the Helicobacter pylori infection group was significantly higher than that in the non-Helicobacter pylori infection group(P<0.05),but after interference by IL-17 blocking antibodies,the expression of LCN2 decreased significantly;similar findings were found in cell experiments that the addition of IL-17 could increase the adhesion of Helicobacter pylori to GES-1 cells and upregulate the expression of LCN2 The addition of IL-17 blocking antibodies can downregulate LCN2 expression.Conclusion:Helicobacter pylori infection may upregulate the expression of the LCN2 gene,which may be regulated by IL-17 cytokines.