RESUMEN
Barrett's esophagus (BE) is a premalignant condition associated with the development of esophageal adenocarcinoma (EAC). Previous studies have implicated hydrophobic bile acids and gastric acid in BE and EAC pathogenesis. In this study, we tested the hypothesis that DNA damage, cytotoxicity and oxidative stress induced by bile acids and gastric acid can be attenuated by the cytoprotective, hydrophilic bile acid glycoursodeoxycholic acid (GUDCA). Non-dysplastic BE cells were exposed for 10 min to pH 4 and/or bile acid cocktail or to pH 4 and a modified cocktail consisting of a mixture of bile acids and GUDCA. DNA damage was evaluated by the comet assay; cell viability and proliferation were measured by trypan blue staining and the MTS assay; reactive oxygen species (ROS) were measured using hydroethidium staining; oxidative DNA/RNA damage was detected by immunostaining with antibody against 8-OH-dG; thiol levels were measured by 5-chloromethylfluorescein diacetate (CMFDA) staining; and the expression of antioxidant proteins was evaluated by western blotting. DNA damage and oxidative stress were significantly increased, while thiol levels were decreased in BE cells treated with pH 4 and bile acid cocktail compared with cells treated with pH 4 alone or untreated cells. Bile acids and low pH also significantly decreased cell proliferation. Expression of the antioxidant enzymes, MnSOD and CuZnSOD, was elevated in the cells treated with bile acids and low pH. When GUDCA was included in the medium, all these effects of pH 4 and bile acids were markedly reduced. In conclusion, treatment of BE cells with acidified medium and a bile acid cocktail at physiologically relevant concentrations induces DNA damage, cytotoxicity, and ROS. The cytoprotective bile acid, GUDCA, inhibits these deleterious effects by inhibiting oxidative stress.
Asunto(s)
Esófago de Barrett/patología , Citoprotección/efectos de los fármacos , Sustancias Protectoras/farmacología , Ácido Ursodesoxicólico/análogos & derivados , 8-Hidroxi-2'-Desoxicoguanosina , Antioxidantes/análisis , Ácidos y Sales Biliares/efectos adversos , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Ácido Desoxicólico/efectos adversos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análisis , Esófago/efectos de los fármacos , Esófago/patología , Depuradores de Radicales Libres/análisis , Ácido Glicoquenodesoxicólico/efectos adversos , Ácido Glicocólico/efectos adversos , Ácido Glicodesoxicólico/efectos adversos , Humanos , Concentración de Iones de Hidrógeno , Estrés Oxidativo/efectos de los fármacos , ARN/efectos de los fármacos , Especies Reactivas de Oxígeno/análisis , Compuestos de Sulfhidrilo/análisis , Superóxido Dismutasa/análisis , Ácido Taurocólico/efectos adversos , Factores de Tiempo , Ácido Ursodesoxicólico/farmacologíaRESUMEN
Bile damage to gastric mucosa may be demonstrated by means of changes in the transmucosal movement of H+ and Na+ ions. In the present study pure 10 mM solutions of taurine and glycine conjugates of chenodeoxycholic acid were instilled into canine Heidenhain pouches. Solutions were prepared at pH 2, 4, and 8, as previous work had shown a greater damaging effect at low pH. The present study confirmed this pH effect, but only with respect to movement of Na+ ion for taurine conjugates. The magnitude of the changes in ionic movements was much greater with pure bile acid solutions than that seen previously with whole bile. These findings are discussed. The greater damage seen below the pKa of the bile acid conjugates suggests that its nonionized form is the more damaging.