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Objective:To explore the effect of bile salt and bile acid on cultured eternalized human gastric mucosa epithelium GES-1 cells. Methods:Cultured eternalized human gastric mucosa epithelium GES-1 cells were treated with media containing 6 different kinds of bile salts and 3 different kinds of bile acids and their mixture with different concentrations: GCDC(glycochenodeoxycholate),GDC(glycodeoxycholate),GC(glycocholate),TCDC(taurochenodeoxycholate),TDC(taurodeoxycholate),TC (taurocholate),LCA(lithocholicacid),CA(cholic acid),DCA(deoxycholic acid)(50μmol/L,250μmol/L,500μmol/L,1000μmol/L),DY(mixture of bile salts) and DS(mixture of bile acids)(250μmol/L,500μmol/L,1000μmol/L,1500μmol/L,2000μmol/L),in comparison with the control group(in normal media without bile salts and bile acids). Cell proliferation was assessed by MTT(3-[4,5-Dimethylthiaolyl]-2,5-diphenyl-tetrazolium bromide) assay for 72 hours with different concentrations and the apoptotic cells were assayed by flow cytometry (FCM) with Annex V-FITC conjugated with propidium iodide(PI) staining for 24 hours with different concentrations(1500,2000μmol/L). Results:There was no significant difference in morphology and cell proliferation in GC group after 24～72 h. Low concentration(50μmol/L) of GCDC,GDC,TCDC,TDC and TC accelerated gastric epithelial cell growth in a dosage-time dependent manner. At middle concentration(250-500μmol/L),it showed positive effect after 24～48 h,while negative effect after 72 h. At high concentration(1000μmol/L),it accelerated gastric epithelial cell growth after 24h and show consistent inhibition even leading to necrosis after 48～72 h. LCA and CA showed a positive effect on the concentration of 50μmol/L after 24～72 h,while 250-1000μmol/L showed a trend towards apoptosis after 24～72 h. At 50-500μmol/L,DCA showed proliferation after 24 h and apoptosis after 48～72 h,but showed necrosis after 24～72 h at 1000 μmol/L. DY and DS could facilitate normal gastric mucosa epithelial cell growth at low concentration (250-500μmol/L),however at 1000-2000μmol/L the trend shifted from apoptosis to necrosis. FCM with Annexin-V conjugated with PI staining revealed that GCDC,GDC,GC,TCDC,TDC,TC,LCA,CA,DCA,DY and DS induced apoptosis of human gastric mucosal epithelial cells. They were all significantly higher than that of the control(P < 0.05),but there was no significant difference in GC group (P > 0.05). The bile salts induced apoptosis in a time-dose-dependent manner. Conclusion:Our results suggested that bile acid and bile salt is the trigger of injury in human gastric mucosal epithelial cells.
Objective: To explore the effect of bile salt and bile acid on cultured eternalized human gastric mucosa epithelium GES-1 cells. Methods: Cultured eternalized human gastric mucosa epithelium GES-1 cells were treated with media containing 6 different kinds of bile salts and 3 different GDC (glycodeoxycholate), GC (glycocholate), TCDC (taurochenodeoxycholate), TDC (taurodeoxycholate), TC (taurocholate), LCA (lithocholic acid), CA ), DCA (250 μmol / L, 500 μmol / L, 1000 μmol / L, 250 μmol / L, 500 μmol / L, 1000 μmol / L) / L, 1500 μmol / L, 2000 μmol / L) in comparison with the control group (in normal media without bile salts and bile acids). Cell proliferation was assessed by MTT (3- [4,5-Dimethylthiaolyl] -diphenyl-tetrazolium bromide assay for 72 hours with different concentrations and the apoptotic cells were assayed by flow cytometr Results: There was no significant difference in morphology and cell proliferation in GC group after 24 ~ 72 (PI) staining for 24 hours with different concentrations (15,000, 2000 μmol / L) (50μmol / L) of GCDC, GDC, TCDC, TDC and TC accelerated gastric epithelial cell growth in a dosage-dependent dependent manner. At middle concentration (250-500μmol / L) 48h, while negative effect after 72 h. At high concentration (1000μmol / L), it accelerated gastric epithelial cell growth after 24h and show consistent inhibition even leading to necrosis after 48 ~ 72 h. LCA and CA showed a positive effect on the concentration of 50μmol / L after 24 ~ 72h, while while 250-1000μmol / L showed a trend toward apoptosis after 24 ~ 72h. At 50-500μmol / L, DCA showed proliferation after 24h and apoptosis after 48 ~ 72h, but showed necrosis after 24 ~ 72 h at 1000 μmol / L. DY and DS could facilitate normal gastric mucos a epithelial cell growth at lowconcentration (250-500 μmol / L) but at 1000-2000 μmol / L the trend shifted from apoptosis to necrosis. FCM with Annexin-V conjugated with PI staining revealed that GCDC, GDC, GC, TCDC, TDC, TC, LCA, CA , DCA, DY and DS induced apoptosis in human gastric mucosal epithelial cells. They were all significantly higher than that of the control (P <0.05), but there was no significant difference in GC group (P> 0.05). The bile salts induced Conclusion: Our results suggested that bile acid and bile salt is the trigger of injury in human gastric mucosal epithelial cells.