Effects of Acute Ethanol Administration on LPS‐Induced Expression of Cyclooxygenase‐2 and Inducible Nitric Oxide Synthase in Rat Alveolar Macrophages

Abstract

Activation of alveolar macrophages acts as a primary defense mechanism of lung with immunologic and inflammatory processes. Incidence of respiratory distress syndrome (ARDS) has been reported to be higher in alcoholics than that in nonalcoholics. Both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are induced by inflammatory stimuli such as LPS and cytokines and are involved in host defense. COX-2 and iNOS have been reported to play important roles in pathophysiology of ARDS. The aim of the present study was to elucidate whether acute ethanol administration to rats affects on COX-2 and iNOS expression in isolated alveolar macrophages. Ethanol (4.5 g per kg body weight as a 20% solution) was intraperitoneally injected to male Wistar rats. At 2.5 hrs after the injection, alveolar macrophages were collected from rats by bronchoalveolar lavage and were stimulated with lipopolysaccharide (LPS, 1 mug/ml). Expression of COX-2 and iNOS and activation of MAPKs was evaluated by Western blotting. In alveolar macrophages isolated from ethanol-treated rats, LPS-stimulated production of both prostaglandin E2 and nitrite was significantly lower than that in macrophages isolated from vehicle-treated control rats. LPS-induced expression of both COX-2 and iNOS was significantly lower in macrophages from ethanol-treated rats than that in macrophages from the control rats, while expression of beta-actin was not different in these groups. LPS increased phosphorylation of both extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein kinase (MAPK). The levels of phosphorylated ERK and p38 were significantly lower in macrophages from ethanol-treated rats compared with those from the control rats. Treatment of macrophages with ethanol (100 - 400 mM) in vitro significantly inhibited expression of COX-2 in a concentration-dependent manner, while only a high concentration (400 mM) of ethanol significantly inhibited expression of iNOS. Ethanol also inhibited COX-2 expression in the presence of Tiron. Expression of COX-2 and iNOS was significantly inhibited by U0126 but not by SB203580. In rat alveolar macrophages, LPS-induced expression of COX-2 and iNOS is mediated by ERK MAPK but not by p38 MAPK. Acute ethanol administration to rats attenuates induction of both COX-2 and iNOS in alveolar macrophages by inhibiting phosphorylation of ERK.