HNE-induced 5-LO expression is regulated by NF-{kappa}B/ERK and Sp1/p38 MAPK pathways via EGF receptor in murine macrophages

5-Lipoxygenase (5-LO) has been suggested to be a modulator of atherosclerotic plaque instability and co-exists with 4-hydroxynonenal (HNE) in macrophages in atherosclerotic lesions. To determine the potential role for HNE in 5-LO expression, the molecular mechanisms of 5-LO expression were evaluated in HNE-stimulated macrophages. A genomic sequence of the promoter 2.0 kb upstream of the transcription initiation site was amplified, and a series of sequentially deleted fragments were then fused to a luciferase reporter gene. The promoter region 213 bp upstream of the transcription start site was responsible for the HNE-enhanced transcriptional activity of 5-LO. Site-directed mutagenesis of this region showed that the transcription factors, including stimulating protein 1 (Sp1) and nuclear factor-κB (NF-κB), were associated with up-regulation of HNE-induced 5-LO transcription. Moreover, the role of Sp1 and NF-κB in HNE-induced 5-LO expression was confirmed by siRNA knockdown of Sp1 and NF-κB. The HNE-enhanced Sp1 and NF-κB activities were attenuated by SB203580, a p38 mitogen-activated protein kinase (MAPK) inhibitor, and PD98059, an extracellular signal-regulated kinase (ERK) inhibitor, respectively. In addition, the HNE-enhanced phosphorylation of p38 MAPK and ERK was inhibited by AG1478, an epidermal growth factor receptor (EGFR) antagonist, but not by AG1295, a platelet-derived growth factor receptor (PDGFR) antagonist. 5-LO expression by HNE was regulated at the transcriptional level by the EGFR-mediated activation of Sp1/p38 MAPK and NF-κB/ERK pathways in macrophages, which may lead to the development of therapeutic interventions for regulating 5-LO expression in atherosclerosis.