Src-dependent STAT-3-mediated Expression of Monocyte Chemoattractant Protein-1 Is Required for 15(S)-Hydroxyeicosatetraenoic Acid-induced Vascular Smooth Muscle Cell Migration

Harihara S.K Potula,Dong Wang,Dong Van Quyen,Nikhlesh K Singh,Venkatesh Kundumani-Sridharan,Manjula Karpurapu,Edwards A Park,Wayne C Glasgow,Gadiparthi N Rao

doi:10.1074/jbc.m109.012526

Abstract

To understand the role of human 15-lipoxygenase 1 (15-LOX1) in vascular wall remodeling, we have studied the effect of the major 15-LOX1 metabolite of arachidonic acid, 15(S)-hydroxyeicosatetraenoic acid (15(S)-HETE), on vascular smooth muscle cell (VSMC) migration both in vitro and in vivo. Among 5(S)-HETE, 12(S)-HETE, and 15(S)-HETE, 15(S)-HETE potentially stimulated more vascular smooth muscle cell (VSMC) migration. In addition, 15(S)-HETE-induced VSMC migration was dependent on Src-mediated activation of signal transducer and activator of transcription-3 (STAT-3). 15(S)-HETE also induced monocyte chemoattractant protein-1 (MCP-1) expression via Src-STAT-3 signaling, and neutralizing anti-MCP-1 antibodies completely negated 15(S)-HETE-induced VSMC migration. Cloning and characterization of a 2.6-kb MCP-1 promoter revealed the presence of four putative STAT-binding sites, and the site that is proximal to the transcription start site was found to be essential for 15(S)-HETE-induced Src-STAT-3-mediated MCP-1 expression. Rat carotid arteries that were subjected to balloon injury and transduced with Ad-15-LOX1 upon exposure to [(3)H]arachidonic acid ex vivo produced 15-HETE as a major eicosanoid and enhanced balloon injury-induced expression of MCP-1 in smooth muscle cells in Src and STAT-3-dependent manner in vivo. Adenovirus-mediated delivery of 15-LOX1 into rat carotid artery also led to recruitment and homing of macrophages to medial region in response to injury. In addition, transduction of Ad-15-LOX1 into arteries enhanced balloon injury-induced smooth muscle cell migration from media to intima and neointima formation. These results show for the first time that 15-LOX1-15(S)-HETE axis plays a major role in vascular wall remodeling after balloon angioplasty.

Highlights

VSMC3 migration from media to intima plays a determinant role in atherosclerosis and restenosis [1,2,3]
Hydroxyeicosatetraenoic Acids Stimulate vascular smooth muscle cell (VSMC) Migration— Previously we have reported that 15(S)-hydroxyeicosatetraenoic acids (HETEs) stimulates VSMC migration in a dose-dependent manner with maximum effect at 0.5 ␮M, which is a pathophysiological concentration [29]
To understand the role of eicosanoids in the pathogenesis of atherosclerosis and restenosis, we first studied the comparative effect of various LOX products of Arachidonic acid (AA), namely, 5(S)-HETE, 12(S)-HETE, and 15(S)-HETE, on VSMC migration using a modified Boyden chamber method

Summary

Introduction

VSMC3 migration from media to intima plays a determinant role in atherosclerosis and restenosis [1,2,3]. Loon injury and transduction with Ad-GFP or Ad-15-LOX1, the a 4-fold increase in MCP-1 promoter-luciferase activity arteries were isolated, and RNA was extracted and analyzed for was observed in response to 15(S)-HETE with pGL3-MCP1(0.2 MCP-1 mRNA levels by RT-PCR.

Results

Conclusion