Role of Endothelin-1 and Constitutive Nitric Oxide Synthase in Gastric Mucosal Resistance to Indomethacin Injury: Effect of Antiulcer Agents

Abstract

Background: Endothelin-1 (ET-1) and nitric oxide, recognized key mediators implicated in the pathophysiology of gastric mucosal injury, are known to exert opposing effects on the inflammatory processes mediated by regulatory cytokines. In this study we investigated the mucosal expression of ET-1 and interleukin-4 (IL-4) and the activity of constitutive nitric oxide synthase (cNOS) during indomethacin-induced gastric mucosal injury and evaluated the effect of antiulcer agents, omeprazole and sucralfate, on this process. Methods: The experiments were conducted with groups of rats pretreated intragastrically with omeprazole (40 mg/kg), sucralfate (200 mg/kg), or vehicle, followed 30 min later by an intragastric dose of indomethacin (60 mg/kg). The animals were killed 2 h later, and their mucosal tissue subjected to macroscopic damage assessment, ET-1 and IL-4 expression assay, and the measurement of cNOS activity. Results: In the absence of antiulcer agents, indomethacin caused multiple hemorrhagic lesions and extensive epithelial cell apoptosis, accompanied by a 20.7% reduction in IL-4, a 3.1-fold increase in mucosal expression of ET-1, and a 4.2-fold decrease in cNOS. Pretreatment with a gastroprotective agent, sucralfate, produced a 59.7% reduction in the mucosal damage caused by indomethacin, a 41.2% decrease in epithelial cell apoptosis, and a 56.5% reduction in ET-1, whereas the expression of IL-4 increased by 29.3% and that of cNOS showed a 3.3-fold increase. In contrast, the pretreatment with a proton pump inhibitor, omeprazole, led to only a 10.5% reduction in the extent of mucosal damage caused by indomethacin and a 13% decrease in apoptosis, whereas the expression of cNOS increased by 68.7% and ET-1 by 12.2%, and the level of IL-4 remained essentially unchanged. Conclusions: The results suggest that an increase in the vasoconstrictive ET-1 level combined with a decrease in regulatory cytokine, IL-4, and a loss of compensatory action by cNOS may be responsible for the gastric mucosal injury caused by indomethacin. Our findings also indicate the value of sucralfate in countering the untoward gastrointestinal side effects of nonsteroidal anti-inflammatory drug therapy.