PPARγ Ligands Regulate NADPH Oxidase, eNOS, and Barrier Function in the Lung Following Chronic Alcohol Ingestion

Background: Chronic alcohol ingestion increases the incidence and severity of the acute respiratory distress syndrome (ARDS), where reactive species contribute to alveolar-capillary barrier dysfunction and noncardiogenic pulmonary edema. Previous studies demonstrated that chronic alcohol ingestion increased lung NADPH oxidase and endothelial nitric oxide synthase (eNOS) expression and that ligands for the peroxisome proliferator-activated receptor gamma (PPAR?) reduced NADPH oxidase expression. Therefore, we hypothesized that the PPAR? ligand, rosiglitazone, would attenuate alcohol-induced NADPH oxidase expression and pulmonary barrier dysfunction. Methods: C57Bl/6 mice were treated +/- alcohol in drinking water (20% w/v) for 12 weeks. During the final week of alcohol treatment, mice were gavaged with rosiglitazone (10 mg/kg/d) or vehicle. Selected animals were treated twice with lipopolysaccharide (LPS, 2 mg/kg IP) prior to sacrifice. Pulmonary barrier dysfunction was estimated from protein content of bronchoalveolar lavage (BAL) fluid. Results: LPS treatment increased BAL protein in alcohol-fed but not control mice, and rosiglitazone attenuated LPS and alcohol-induced pulmonary barrier dysfunction. Alcohol- and LPS-induced increases in lung eNOS, Nox1, and Nox4 expression were attenuated by rosiglitazone. In vitro, alcohol (0.10% w/v) increased H2O2 production, barrier dysfunction, eNOS, Nox1, and Nox4 expression in human umbilical vein endothelial cell (HUVEC) monolayers, effects also attenuated by rosiglitazone (10 mu M). Alcohol-induced HUVEC barrier dysfunction was attenuated by inhibition of NOS or addition of the eNOS cofactor, tetrahydrobiopterin. Conclusions: These results indicate that PPAR gamma activation reduced expression of eNOS, Nox1, Nox4, the production of reactive species, and barrier dysfunction caused by chronic alcohol ingestion and suggest that PPAR gamma represents a novel therapeutic target for strategies designed to reduce the risk of lung injury in patients with a history of chronic alcohol ingestion.