Expression Of S100A8/A9 In Human Gastric Epithelial GES-1 Cells Induced By Helicobacter Pylori

Objective:Establish a model of human gastric epithelial GES-1 cells infected with Helicobacter pylori(H.pylori)in vitro.To investigate the effect of H.pylori on S100A8,S100A9 in human gastric epithelial GES-1 cells and the relationship between the level of S100A8 and S100A9 and the degree of inflammation induced by H.pylori in GES-1 cells.Methods:The H.pylori isolates 26995 were co-cultured with human gastric epithelial cells GES-1 cells for 24 h at different multiplicity of infection(MOI of 0,50:1,100:1 and 200:1 respectively).The proliferation activity of GES-1 cells was detected by CCK-8 method.The expression of S100A8 and S100A9 m RNA was detected by RT-PCR.The expression of S100A8,S100A9 and inflammatory cytokines TNF-α was detected by ELISA method.Then analyzed the correlation between S100A8,S100A9 protein and inflammatory factor expression.Results:1.Compared with the blank control group,after 24 h co-culture with H.pylori,GES-1 had obvious morphological changes.The specific manifestation was that the cells became round and swollen,the polarity disappeared,and the cytoplasm was vacuolized.The number of suspended cells began to increase.2.Compared with the blank control group,the proliferation activity of GES-1 cells was increased when co-cultured with H.pylori at the low infection multiplicity(MOI was50:1,100:1),while the proliferation activity of the GES-1 cells is reduced when the infection multiplicity(MOI was 200:1)was high.3.Compared with the blank control group,the level of inflammatory cytokines TNF-α in the culture supernatant of GES-1 cells co-cultured with H.pylori for 24 hours was increased,and the difference between the experimental group and the control group was statistically significant.4.Compared with the blank control group,the expression of S100A8 and S100A9 m RNA in H.pylori-infected GES-1 cells all increased to some extent,and the S100A8 and S100A9 proteins in the supernatant of the cell culture also increased to some extent,The difference was statistically significant.5.In the different infection multiplicity H.pylori infection GES-1 cell model,S100A8,S100A9 protein expression in cell culture supernatant was positively correlated with the level of inflammatory factor TNF-α.Conclusion:1.Co-cultured with H.pylori for 24 h,there were morphological changes,proliferative activity disorder and secretion of inflammatory factors in GES-1 cells,indicating that infection in vitro GES-1 cell infected with H.pylori model in vitro was successfully established;2.H.pylori could induce the high expression of S100A8 and S100A9 m RNA in human gastric epithelial GES-1 cells and increase the secretion of S100A8 and S100A9 proteins.3.S100A8,S100A9 can reflect the infection of H.pylori GES-1 cells in the degree of inflammationto to some extent.