Viral vector-mediated 12/15-lipoxygenase overexpression in vascular smooth muscle cells enhances inflammatory gene expression and migration
Viral vector-mediated 12/15-lipoxygenase overexpression in vascular smooth muscle cells enhances inflammatory gene expression and migration
复制标题
病毒载体介导的血管平滑肌细胞中12/15-脂氧合酶过度表达增强炎症基因表达和迁移
DOI：
10.1159/000109966
复制
发表时间：
2008-01-01
影响因子：
1.7
通讯作者：
Reddy, Marpadga A.
中科院分区：
文献类型：
--
作者：
Dwarakanath, Roopashree S.;Sahar, Saurabh;Reddy, Marpadga A.
血管平滑肌细胞(VSMCs)上12/15-脂氧合酶(12/15-LO)的表达和活性增加在糖尿病和血管并发症的发病机制中起关键作用。然而，12/15-LO过表达对VSMC迁移和炎性基因表达的影响尚不清楚。在这项研究中，12/15-LO经腺病毒和杆状病毒载体在人VSMC(HVSMCs)中过表达，并与对照增强型绿色荧光蛋白(EGFP)表达的细胞进行了动脉粥样硬化反应的比较。转导12/15-LO病毒的人血管内皮细胞表达高水平的酶活性蛋白，并产生较高水平的LO产物--12(S)-羟基二十碳四烯酸。12/15-LO过表达的人肾小管上皮细胞与表达绿色荧光蛋白的人肾小管上皮细胞相比，氧化应激增加、p38丝裂原活化蛋白激酶活化、迁移和炎症基因表达增加。此外，抗氧化剂、针对p65(核因子-kappa B)的siRNAs或表达抑制性I-kappa Bα或I-kappa B超阻遏突变体的新一代杆状病毒可取消12/15-LO过表达所诱导的炎症基因表达。因此，我们首次使用包括杆状病毒在内的新型病毒载体递送系统将外源基因转移到VSMC中，从而证明了12/15-LO的过表达增加了VSMC中的氧化应激、丝裂原激活的蛋白激酶激活、迁移和炎症基因，并且NF-kappa B是一个关键的下游效应因子。病理条件下12/15-LO水平升高引起的VSMCs致动脉粥样硬化反应增强可能是导致血管功能障碍的原因之一。版权所有(C)2007 S.Karger AG，巴塞尔。
Increased expression and activity of 12/15-lipoxygenase (12/15-LO) in vascular smooth muscle cells (VSMCs) play a key role in the pathogenesis of diabetes and vascular complications. However, the consequences of 12/15-LO overexpression for VSMC migration and inflammatory gene expression are not known. In this study, 12/15-LO was over-expressed using adeno- and baculoviral vectors in human VSMC (HVSMCs) and proatherogenic responses compared with control enhanced green fluorescent protein (EGFP)-expressing cells. HVSMCs transduced with 12/15-LO viruses expressed high levels of enzymatically active protein and produced increased levels of the LO product, 12( S)- hydroxyeicosatetraenoic acid. 12/15-LO-overexpressing HVSMCs exhibited increased oxidant stress, activation of p38 mitogen-activated protein kinase, migration and inflammatory gene expression relative to HVSMCs expressing EGFP. Furthermore, inflammatory gene expression induced by 12/15-LO overexpression was abolished by anti-oxidants, siRNAs targeting p65 (nuclear factor-kappa B), or new-generation baculoviruses expressing inhibitory I kappa B alpha or I kappa B superrepressor mutant. Thus, we have used novel viral vector delivery systems, including baculoviruses, for the first time to deliver foreign genes into VSMCs and thereby demonstrated that 12/15-LO overexpression increases oxidant stress, mitogen-activated protein kinase activation, migration and inflammatory genes in VSMCs and that NF-kappa B is a key downstream effector. Enhanced proatherogenic responses in VSMCs triggered by increased 12/15-LO levels under pathological conditions may contribute to vascular dysfunction. Copyright (C) 2007 S. Karger AG, Basel.