γ-Glutamyl transpeptidase (GGT) plays key roles in the metabolism of glutathione. Previous studies have shown that GGT expression was increased by oxidants, but the mechanism remains unclear. In the present study, the effects of 4-hydroxy-2-nonenal (HNE), an electrophilic end product of lipid peroxidation, on GGT expression were investigated in rat lung epithelial type II (L2) cells. We demonstrated that HNE increased GGT activity and mRNA content in both time- and dose-dependent manners. Actinomycin D, an RNA transcription inhibitor, blocked HNE-stimulated increase in GGT mRNA, suggesting transcriptional regulation of GGT mRNA by HNE. Of the seven GGT mRNA transcripts known to be produced from the single rat GGT gene, we found that types I, II, and V-2 were constitutively expressed in L2 cells, but only types I and V-2 were increased by HNE. PD98059 and SB203580, relatively specific inhibitors of the ERK and the p38MAPK kinase pathway, respectively, significantly attenuated HNE induction of both GGT activity and mRNA content. In contrast, studies with JNK inhibitor I, a cell-permeable peptide, indicated that JNK was not involved in the GGT induction by HNE. We also found that GGT induction by HNE could be completely blocked by a cocktail of PD98059 and SB203580, suggesting a combined effect of ERK and p38MAPK pathways in HNE-mediated GGT induction. In conclusion, our results demonstrate that HNE increased GGT expression in rat alveolar type II cells and that the induction of GGT by HNE was mediated through activation of the ERK and p38MAPK pathways. © 2004 Elsevier Inc. All rights reserved.