Oxidation modifies the structure and function of the extracellular matrix generated by human coronary artery endothelial cells
Oxidation modifies the structure and function of the extracellular matrix generated by human coronary artery endothelial cells
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DOI：
10.1042/bj20131471
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发表时间：
2014-04-15
影响因子：
4.1
通讯作者：
Davies, Michael J.
中科院分区：
文献类型：
--
作者：
Chuang, Christine Y.;Degendorfer, Georg;Davies, Michael J.
ECM（细胞外基质）材料，如层粘连蛋白、perlecan、IV型胶原和纤维连接蛋白，在决定动脉壁的结构和与ECM相互作用的细胞的特性方面起着关键作用。本研究旨在探讨活化巨噬细胞产生的氧化剂过氧亚硝酸对人冠状动脉内皮细胞（HCAECs）体外和体内ECM结构和功能的影响。我们发现，将hcaec衍生的天然基质成分暴露于浓度为100 μ M的过氧亚硝酸（但不是分解的氧化剂）中，会导致对perlecan、胶原IV以及层粘连蛋白和纤维连接蛋白上的细胞结合位点的抗体识别丧失。识别丧失伴随着HCAEC粘连下降。Real-time PCR结果显示，在过氧亚硝酸处理的hcaec中，与天然基质相比，炎症相关基因MMP7 （matrix metalloproteinase 7）和MMP13上调，层粘连蛋白α 2链下调。免疫组织化学研究证明，在II-III/IV型人类动脉粥样硬化病变中，层粘连蛋白与3-硝基酪氨酸（一种过氧亚硝酸损伤的生物标志物）共定位，与体内疾病发展过程中发生的基质损伤一致。本研究的结果表明，在动脉粥样硬化病变中，过氧亚硝酸改变内皮细胞来源的动脉基底膜天然ECM蛋白的机制。ECM的这些变化，特别是perlecan和层粘连蛋白的变化，可能在诱导细胞功能障碍和促进动脉粥样硬化中起重要作用。
ECM (extracellular matrix) materials, such as laminin, perlecan, type IV collagen and fibronectin, play a key role in determining the structure of the arterial wall and the properties of cells that interact with the ECM. The aim of the present study was to investigate the effect of peroxynitrous acid, an oxidant generated by activated macrophages, on the structure and function of the ECM laid down by HCAECs (human coronary artery endothelial cells) in vitro and in vivo. We show that exposure of HCAEC-derived native matrix components to peroxynitrous acid (but not decomposed oxidant) at concentrations >1 mu M results in a loss of antibody recognition of perlecan, collagen IV, and cell-binding sites on laminin and fibronectin. Loss of recognition was accompanied by decreased HCAEC adhesion. Real-time PCR showed up-regulation of inflammation-associated genes, including MMP7 (matrix metalloproteinase 7) and MMP13, as well as down-regulation of the laminin alpha 2 chain, in HCAECs cultured on peroxynitrous acid-treated matrix compared with native matrix. Immunohistochemical studies provided evidence of co-localization of laminin with 3-nitrotyrosine, a biomarker of peroxynitrous acid damage, in type II-III/IV human atherosclerotic lesions, consistent with matrix damage occurring during disease development in vivo. The results of the present study suggest a mechanism through which peroxynitrous acid modifies endothelial cell-derived native ECM proteins of the arterial basement membrane in atherosclerotic lesions. These changes to ECM and particularly perlecan and laminin may be important in inducing cellular dysfunction and contribute to atherogenesis.