Abstract

Pro-oxidative and pro-inflammatory pathways in vascular endothelium have been implicated in the critical early pathological events in atherosclerosis. The present study was designed to investigate the role of 5-lipoxygenase (5-LOX) in IL-4-induced reactive oxygen species (ROS) generation and MCP-1 expression in vascular endothelium. Real-time RT-PCR and ELISA showed that IL-4 significantly up-regulated the mRNA and protein expression of MCP-1 in primary human aortic endothelial cells (HAEC) and mouse aortas. To determine whether 5-LOX is involved in IL-4-induced MCP-1 expression, HAEC were pre-treated with nordihydroguaiaretic acid (NDGA), a selective 5-LOX inhibitor, for 30 min and then exposed to 10 ng/ml of IL-4. Inhibition of 5-LOX by NDGA significantly attenuated IL-4-induced overexpression of MCP-1 mRNA and protein in HAEC. Additionally, dihydroethidium (DHE) and dichlorodihydrofluorescein (DCF) fluorescence staining showed that pre-treatment of HAEC with NDGA significantly suppressed IL-4-induced ROS generation. Our results suggest that IL-4 induces ROS generation and MCP-1 expression in vascular endothelium through 5-LOX-dependent pathway. These data may contribute to understanding the cellular and molecular mechanisms involved in IL-4-mediated progression of atherosclerosis. (This work was supported by NIH/NHLBI [Grant#R15HL085229] and NSF MILES IGERT).

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