Homocysteine induces VCAM-1 gene expression through NF-κB and NAD(P)H oxidase activation:: protective role of Mediterranean diet polyphenolic antioxidants
Homocysteine induces VCAM-1 gene expression through NF-κB and NAD(P)H oxidase activation:: protective role of Mediterranean diet polyphenolic antioxidants
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DOI：
10.1152/ajpheart.00432.2007
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发表时间：
2007-10-01
影响因子：
4.8
通讯作者：
De Caterina, Raffaele
中科院分区：
文献类型：
--
作者：
Carluccio, Maria Annunziata;Ancora, Maria Assunta;De Caterina, Raffaele
高同型半胱氨酸血症是公认的血管疾病危险因素，但其血管作用的发病机制在很大程度上尚不清楚。由于 VCAM-1 表达在单核细胞粘附和早期动脉粥样硬化形成中至关重要，因此我们评估了同型半胱氨酸 (Hcy) 与 NF-κ B 相关的 VCAM-1 诱导作用，以及膳食多酚抗氧化剂（例如反式白藜芦醇 (RSV) 和羟基酪醇 (HT)）可能的抑制作用，它们是已知的动脉粥样硬化抑制剂 NF-κ B 介导的 VCAM-1 诱导。在人脐静脉内皮细胞 (HUVEC) 中，酶联免疫分析和 Northern 分析分别显示，100 μmol/l 的 Hcy（而非半胱氨酸）在蛋白质和 mRNA 水平上诱导 VCAM-1 表达。使用缺失型 VCAM-1 启动子构建体的转染研究表明，VCAM-1 启动子中的两个串联 NF-κ B 基序对于 Hcy 诱导的 VCAM-1 基因表达是必需的。 EMSA 证实了 Hcy 诱导的 NF-kappa B 激活，Western 分析显示其 p65 (RelA) 亚基的核易位以及抑制剂 I kappa B-alpha 和 I kappa B-beta 的降解。 Hcy 还通过 NAD(P) H 氧化酶激活增加细胞内活性氧，如其 p47(phox) 亚基的膜易位所示。 NF-κ B 抑制剂降低 Hcy 诱导的细胞内活性氧和 VCAM-1 表达。最后，我们发现与营养相关的 RSV 和 HT 浓度（但不包括叶酸和维生素 B6）可减少（在 10(-6) mol/l 时减少 >60%）Hcy 诱导的 VCAM-1 表达和单核细胞对内皮的粘附。这些数据表明，病理生理学相关的 Hcy 浓度通过涉及 NF-kappa B 的促氧化机制诱导 VCAM-1 表达。天然地中海饮食抗氧化剂可以抑制这种激活，表明它们在 Hcy 诱导的血管损伤中可能发挥治疗作用。
Hyperhomocysteinemia is a recognized risk factor for vascular disease, but pathogenetic mechanisms involved in its vascular actions are largely unknown. Because VCAM-1 expression is crucial in monocyte adhesion and early atherogenesis, we evaluated the NF-kappa B-related induction of VCAM-1 by homocysteine (Hcy) and the possible inhibitory effect of dietary polyphenolic antioxidants, such as trans-resveratrol (RSV) and hydroxytyrosol (HT), which are known inhibitors of NF-kappa B-mediated VCAM-1 induction. In human umbilical vein endothelial cells (HUVEC), Hcy, at 100 mu mol/l, but not cysteine, induced VCAM-1 expression at the protein and mRNA levels, as shown by enzyme immunoassay and Northern analysis, respectively. Transfection studies with deletional VCAM-1 promoter constructs demonstrated that the two tandem NF-kappa B motifs in the VCAM-1 promoter are necessary for Hcy-induced VCAM-1 gene expression. Hcy-induced NF-kappa B activation was confirmed by EMSA, as shown by the nuclear translocation of its p65 (RelA) subunit and the degradation of the inhibitors I kappa B-alpha and I kappa B-beta by Western analysis. Hcy also increased intracellular reactive oxygen species by NAD(P) H oxidase activation, as shown by the membrane translocation of its p47(phox) subunit. NF-kappa B inhibitors decreased Hcy-induced intracellular reactive oxygen species and VCAM-1 expression. Finally, we found that nutritionally relevant concentrations of RSV and HT, but not folate and vitamin B6, reduce (by >60% at 10(-6) mol/l) Hcy-induced VCAM-1 expression and monocytoid cell adhesion to the endothelium. These data indicate that pathophysiologically relevant Hcy concentrations induce VCAM-1 expression through a prooxidant mechanism involving NF-kappa B. Natural Mediterranean diet antioxidants can inhibit such activation, suggesting their possible therapeutic role in Hcy-induced vascular damage.