Role of Endothelin-converting Enzyme-1 in the Suppression of Constitutive Nitric Oxide Synthase in Rat Gastric Mucosal Injury by Indomethacin

Abstract

Background: Disturbances in nitric oxide generation and the release of a vasoactive peptide, endothelin-1 (ET-1), are well recognized early events in pathogenesis of NSAID-induced gastropathy. In this study using phosphoramidon, a potent inhibitor of endothelin-converting enzyme-1 (ECE-1), we investigated the influence of ET-1 on the expression of constitutive (cNOS) and inducible nitric oxide synthase (NOS-2) during gastric mucosal injury caused by indomethacin. Methods: The experiments were conducted with groups of rats pretreated intragastrically with phosphoramidon (10, 20, and 40 mg/kg) or vehicle, followed 30 min later by an intragastric dose of indomethacin (60 mg/kg). The animals were killed 4 h later and their mucosal tissue subjected to macroscopic damage assessment and biochemical measurements. Results: In the absence of phosphoramidon, indomethacin caused extensive multiple hemorrhagic lesions of glandular mucosa, accompanied by a 29.9-fold increase in epithelial cell apoptosis, a 13.3-fold increase in NOS-2 and a 5.5-fold decline in the activity of cNOS, while the mucosal expression of ECE-1 activity increased 4-fold and the level of ET-1 showed a 4.8-fold increase. Pretreatment with phosphoramidon produced dose-dependent reduction in the extent of mucosal damage caused by indomethacin, accompanied by a significant recovery in the expression of cNOS, and a marked decline in ECE-1, epithelial cell apoptosis and the mucosal level of ET-1. Phosphoramidon, however, had no effect on the indomethacin-induced increase in the mucosal expression of NOS-2. Conclusions: The results suggest that suppression of ET-1 generation counters the mucosal drop in cNOS and the extent of gastric mucosal damage caused by indomethacin, but has no effect on the mucosal responses associated with up-regulation of NOS-2 expression. Hence, only cNOS plays a role in the protection of gastric mucosa against damage by NSAIDs.