Ethanol (EtOH)-Induced TGF-ß1 and Reactive Oxygen Species Production Are Necessary for EtOH-Induced Alveolar Macrophage Dysfunction and Induction of Alternative Activation

Background Previous studies have shown that chronic ethanol (EtOH) ingestion results in impaired alveolar macrophage function, increased TGF-beta 1 production, and decreased antioxidant availability. Similarly, alternative activation (M2 activation) of alveolar macrophages also induces TGF-beta 1 production and impairs macrophage function. However, the potential links between EtOH-induced alveolar macrophage derangements, M2 activation, TGF-beta 1 production signaling, and oxidant stress have yet to be examined. We hypothesized that EtOH-induced oxidant stress and induction of TGF-beta 1 signaling result in alternative activation which subsequently impairs the phagocytic capacity of alveolar macrophages. Methods Primary rat alveolar macrophages and the alveolar macrophages cell line NR8383 were treated with 0.08% EtOH +/- the antioxidant glutathione (GSH) or a TGF-beta 1 neutralizing antibody for 5 days. Outcome measures included TGF-beta 1 production, reactive oxygen species (ROS) production, phagocytic capacity, and expression of markers of M2 activation. Results Chronic EtOH treatment greatly decreased alveolar macrophage phagocytic function, increased ROS production, increased TGF-beta 1, and increased expression of markers of M2 activation. GSH supplementation and inhibition of TGF-beta 1 signaling during EtOH treatment prevented these alterations. Conclusions EtOH treatment increased oxidant stress, TGF-beta 1 production, and alternative activation in NR8383 cells. However, GSH supplementation and ablation of TGF-beta 1 signaling prevented these effects. This suggested that the EtOH-induced switch to an M2 phenotype was a result of decreased antioxidant availability and increased TGF-beta 1 signaling. Preventing EtOH-induced induction of alternative activation may improve alveolar macrophage function in alcoholic subjects and decrease the risk of respiratory infections.