CREB-mediated IL-6 expression is required for 15(S)-hydroxyeicosatetraenoic acid-induced vascular smooth muscle cell migration.
CREB-mediated IL-6 expression is required for 15(S)-hydroxyeicosatetraenoic acid-induced vascular smooth muscle cell migration.
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DOI：
10.1161/atvbaha.109.185777
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发表时间：
2009-06
期刊：
影响因子：
--
通讯作者：
Rao GN
中科院分区：
文献类型：
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作者：
Chava KR;Karpurapu M;Wang D;Bhanoori M;Kundumani-Sridharan V;Zhang Q;Ichiki T;Glasgow WC;Rao GN
血管平滑肌细胞（VSMCs）从中膜向内膜的迁移是动脉粥样硬化和再狭窄病理生理学中的关键事件。多不饱和脂肪酸（PUFA）的脂氧合酶产物被证明在这些疾病中发挥作用。环磷酸腺苷反应元件结合蛋白（CREB）参与了凝血酶和血管紧张素II对VSMC生长和运动的调节。本研究的目的是检测CREB在氧化脂质分子15（S）-HETE诱导的VSMC迁移和新生内膜形成中的作用。15（S）-HETE以CREB依赖的方式刺激VSMC迁移，如通过改良的Boyden室法测量的。阻断MEK 1、JNK 1或p38 MAPK可抑制15（S）-HETE诱导的CREB磷酸化和VSMC迁移。15（S）-HETE诱导白细胞介素-6（IL-6）的表达和分泌，分别通过RT-PCR和ELISA分析。中和抗IL-6抗体阻断15（S）-HETE诱导的VSMC迁移。显性负性MUR介导的ERK 1/2、JNK 1、p38 MAPK或CREB阻断可抑制15（S）-HETE诱导的VSMC中IL-6的表达。IL-6启动子的连续5′端缺失和沿着的定点突变以及使用抗CREB抗体的染色质免疫沉淀显示cAMP反应元件对于15（S）-HETE诱导的IL-6表达是必需的。显性负性CREB还抑制球囊损伤诱导的IL-6表达、SMC从中膜向内膜区迁移和新生内膜形成。腺病毒介导的15-脂氧合酶2（15-LOX 2）转导引起VSMCs中15-HETE的产生增加，并增强IL-6的表达、SMC从中膜向内膜区的迁移以及对动脉损伤的响应的新生内膜形成。上述结果表明15-LOX 2 -15-HETE在涉及CREB介导的IL-6表达的VSMC迁移和新生内膜形成的调节中的作用。
Migration of vascular smooth muscle cells (VSMCs) from media to intima is a key event in the pathophysiology of atherosclerosis and restenosis. The lipoxygenase products of polyunsaturated fatty acids (PUFA) were shown to play a role in these diseases. Cyclic AMP response element binding protein (CREB) has been implicated in the regulation of VSMC growth and motility in response to thrombin and angiotensin II. The aim of the present study was to test the role of CREB in an oxidized lipid molecule, 15(S)-HETE-induced VSMC migration and neointima formation. 15(S)-HETE stimulated VSMC migration in CREB-dependent manner, as measured by the modified Boyden chamber method. Blockade of MEK1, JNK1 or p38MAPK inhibited 15(S)-HETE-induced CREB phosphorylation and VSMC migration. 15(S)-HETE induced expression and secretion of interleukin-6 (IL-6), as analyzed by RT-PCR and ELISA, respectively. Neutralizing anti-IL-6 antibodies blocked 15(S)-HETE-induced VSMC migration. Dominant-negative mutant-mediated blockade of ERK1/2, JNK1, p38MAPK or CREB suppressed 15(S)-HETE-induced IL-6 expression in VSMCs. Serial 5′ deletions and site-directed mutagenesis of IL-6 promoter along with chromatin immunoprecipitation using anti-CREB antibodies showed that cAMP response element is essential for 15(S)-HETE-induced IL-6 expression. Dominant-negative CREB also suppressed balloon injury-induced IL-6 expression, SMC migration from media to intimal region and neointima formation. Adenovirus-mediated transduction of 15-lipoxygenase 2 (15-LOX2) caused increased production of 15-HETE in VSMCs and enhanced IL-6 expression, SMC migration from media to intimal region and neointima formation in response to arterial injury. The above results suggest a role for 15-LOX2-15-HETE in the regulation of VSMC migration and neointima formation involving CREB-mediated IL-6 expression.