Ethanol renders the lung susceptible to acute lung injury in the setting of insults such as sepsis. The mechanisms mediating this effect are unknown, but activation of tissue remodeling is considered key to this process. We found that chronic ethanol ingestion in rats increased the expression of fibronectin, a matrix glycoprotein implicated in acute lung injury. In cultured NIH/3T3 cells and in primary rat and mouse lung fibroblasts, ethanol induced fibronectin mRNA and protein expression in a dose- and time-dependent fashion. The effect of ethanol was prevented by inhibitors of protein kinase C and mitogen-activated protein kinases, and was associated with the phosphorylation and increased DNA binding of the transcription factor CREB followed by increased transcription of the fibronectin gene. Fibroblasts were found to express 7 nicotinic acetylcholine receptor (nAChR) and ethanol induction of fibronectin was abolished by -bungarotoxin and methyllcaconitine, inhibitors of 7 nAChRs. However, ethanol was able to induce fibronectin mRNA and protein in primary lung fibroblasts isolated from 7 nicotinic acetylcholine receptor knockout mice. The ethanol-induced fibronectin response was dependent on ethanol metabolism since 4-methypyrazole, an inhibitor of alcohol dehydrogenase, abolished the effect and acetaldehyde induced it. These observations suggest that ethanol or ethanol metabolites stimulate lung fibroblasts to produce fibronectin by inducing specific signals transmitted via nAChRs independent of the 7 subunit, and this might represent a mechanism by which ethanol renders the lung susceptible to acute lung injury.