Cytochrome p450 (CYP) 2J2 gene transfection attenuates MMP-9 via inhibition of NF-κβ in hyperhomocysteinemia
Cytochrome p450 (CYP) 2J2 gene transfection attenuates MMP-9 via inhibition of NF-κβ in hyperhomocysteinemia
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DOI：
10.1002/jcp.21356
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发表时间：
2008-06-01
影响因子：
5.6
通讯作者：
Tyagi, Suresh C.
中科院分区：
文献类型：
--
作者：
Mosha, Karni S.;Zeldin, Darryl C.;Tyagi, Suresh C.
高同型半胱氨酸血症（HHcy）与花生四烯酸（AA）代谢调节和基质金属蛋白酶-9 （MMP-9）激活相关的动脉粥样硬化事件。细胞色素P450 （CYP）环氧化酶2j2 （CYP2J2）在心脏内皮中含量丰富，其AA代谢物环氧二碳三烯酸（EETs）通过nf - κ β减轻炎症。然而，CYP2J2在HHcy中调控MMP-9的潜在分子机制尚不清楚。我们试图确定P450环氧化酶基因转染或EETs补充减弱同型半胱氨酸（Hcy）诱导的MMP-9激活的分子机制。通过pcDNA3.1/CYP2J2载体转染小鼠主动脉内皮细胞（MAECs），发现CYP2J2过表达。转染P450环氧合酶或外源补充EETs对NF-kappa β介导的MMP-9调节的影响采用Western blot、凝胶内明胶酶谱法、电迁移位移法和免疫细胞化学进行评估。结果表明，Hcy下调CYP2J2蛋白表达，使pi3k依赖性AKT信号去磷酸化。Hcy通过下调IK β α （NF-kappa β的内源性胞质抑制剂）诱导NF-kappa β的核易位。Hcy通过增加NF-kappa β - dna结合诱导MMP-9活化。此外，P450环氧化酶转染或外源性添加8,9- eet可使AKT磷酸化，并减弱hcy诱导的MMP-9活化。这在一定程度上是由于NF-kappa β核易位、NF-kappa β dna结合和IK β α活化受到抑制。该研究明确表明eet在Hcy/MMP-9信号调制中的关键作用。
Hyperhomocysteinemia (HHcy) is associated with atherosclerotic events involving the modulation of arachidonic acid (AA) metabolism and the activation of matrix metalloproteinase-9 (MMP-9). Cytochrome P450 (CYP) epoxygenase-2J2 (CYP2J2) is abundant in the heart endothelium, and its AA metabolites epoxyeicosatrienoic acids (EETs) mitigates inflammation through NF-kappa beta. However, the underlying molecular mechanisms for MMP-9 regulation by CYP2J2 in HHcy remain obscure. We sought to determine the molecular mechanisms by which P450 epoxygerase gene transfection or EETs supplementation attenuate homocysteine (Hcy)-induced MMP-9 activation. CYP2J2 was over-expressed in mouse aortic endothelial cells (MAECs) by transfection with the pcDNA3.1/CYP2J2 vector. The effects of P450 epoxygenase transfection or exogenous supplementation of EETs on NF-kappa beta-mediated MMP-9 regulation were evaluated using Western blot, in-gel gelatin zymography, electromobility shift assay, immunocytochemistry. The result suggested that Hcy downregulated CYP2J2 protein expression and dephosphorylated PI3K-dependent AKT signal. Hcy induced the nuclear translocation of NF-kappa beta via downregulation of IK beta alpha (endogenous cytoplasmic inhibitor of NF-kappa beta). Hcy induced MMP-9 activation by increasing NF-kappa beta-DNA binding. Moreover, P450 epoxygerase transfection or exogenous addition of 8,9-EET phosphorylated the AKT and attenuated Hcy-induced MMP-9 activation. This occurred, in part, by the inhibition of NF-kappa beta nuclear translocation, NF-kappa beta-DNA binding and activation of IK beta alpha. The study unequivocally suggested the pivotal role of EETs in the modulation of Hcy/MMP-9 signal.