ROLE OF VEGF IN ATHEROGENESIS

VEGF 在动脉粥样硬化形成中的作用

• 批准号: 10671061
• 负责人: KUZUYA Masafumi
• 金额: $ 1.92万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671061/
• 关键词: atherosclerosis; vascular endothelial cell; vascular endothelial growth factor; oxidized low density lipoprotein; macrophage; VEGF receptor; glutathione; angiogenesis; 変性低比重リポ蛋白

Immunohistochemical studies showed that human early atherosclerotic lesions exhibited intensive vascular endothelial growth factor (VEGF) immunoreactivity in subendothelial macrophage-rich regions of thickened intima. In atherosclerotic plaques, VEGF staining was also observed in foam cell-rich regions adjacent to lipid core or neovascularized basal regions of plaque consisting predominantly of smooth muscle cells. High powered field observation revealed that VEGF was localized in extracellular space as well as macrophage cell surface. These observations suggest the possible involvement of oxidized loe density lipoprtein (Ox-LDL) in the development of human atherosclerosis through VEGF induction in macrophages. We also demonstrated that Ox-LDL upregulated VEGF mRNA expression in RAW 264 cells, monocytic cell line, in a time and concentration dependent manner, and that Ox-LDL stimulated the VEGF protein secretion from the cells.We hypothesized that VEGF in subendothelial macrophage-rich … More regions adjacent to endothelial cells at the luminal surface may participate in the maintenance and repair of the lumen endothelium. To test our hypothesis we examined whether VEGF protects the toxicity of Ox-LDL to cultured endothelial cells derived from bovine aorta (BAECs). BAECs preincubation with VEGF prevented Ox-LDL toxicity in a preincubation time- and VEGF concentration-dependent manner. Incubation of BAECs with VEGF increased intracellular glutathione (GSH) in a time-dependent manner. Combined addition of VEGF and L-buthionine sulfoximine, a GSH synthesis inhibitor, reversed GSH level and the protective effect of VEGF on Ox-LDL-induced cytotoxicity. Placenta growth factor, which ligates to VEGF Fit-1 receptor but not KDR/Flk-1, failed to prevent Ox-LDL toxicity and had no effect on intracellular GSH level. Anti- KDR/Flk-1 antibody completely blocked these activities of VEGF.These results suggest that VEGF prevents Ox-LDL-induced endothelial cell damage via intracellular GSH-dependent mechanism through KDR/Flk-1 receptor. Less

免疫组织化学研究表明，人类早期动脉粥样硬化病变在增厚内膜的内皮下巨噬细胞丰富区域表现出强烈的血管内皮生长因子（VEGF）免疫反应性。在动脉粥样硬化斑块中，在邻近脂质核心的富含泡沫细胞的区域或主要由平滑肌细胞组成的斑块的新生血管基底区域中也观察到 VEGF 染色。高倍视野观察显示VEGF定位于细胞外空间以及巨噬细胞表面。这些观察结果表明，氧化洛伊密度脂蛋白（Ox-LDL）可能通过巨噬细胞中的 VEGF 诱导参与人类动脉粥样硬化的发展。我们还证明，Ox-LDL 以时间和浓度依赖性方式上调 RAW 264 细胞（单核细胞系）中 VEGF mRNA 的表达，并且 Ox-LDL 刺激细胞分泌 VEGF 蛋白。我们假设，在管腔表面邻近内皮细胞的富含内皮下巨噬细胞的区域中的 VEGF 可能参与管腔内皮的维护和修复。为了检验我们的假设，我们检查了 VEGF 是否可以保护 Ox-LDL 对培养的牛主动脉内皮细胞 (BAEC) 的毒性。 BAECs 与 VEGF 预孵育以预孵育时间和 VEGF 浓度依赖性方式防止 Ox-LDL 毒性。 BAEC 与 VEGF 一起孵育会以时间依赖性方式增加细胞内谷胱甘肽 (GSH)。联合添加 VEGF 和 L-丁硫氨酸亚砜亚胺（一种 GSH 合成抑制剂）可逆转 GSH 水平以及 VEGF 对 Ox-LDL 诱导的细胞毒性的保护作用。胎盘生长因子与 VEGF Fit-1 受体连接，但不与 KDR/Flk-1 连接，无法预防 Ox-LDL 毒性，并且对细胞内 GSH 水平没有影响。抗KDR/Flk-1抗体完全阻断了VEGF的这些活性。这些结果表明VEGF通过KDR/Flk-1受体通过细胞内GSH依赖性机制防止Ox-LDL诱导的内皮细胞损伤。较少的

项目成果

Nakayama Shinsuke: "The alpha 1-subunit of smooth muscle Ca^<2+> channel preserves multiple open states induced by depolaruzation."J.of Physiol.. 526. 47-56 (2000)

Nakayama Shinsuke：“平滑肌 Ca^2 通道的 α1 亚基保留了去极化诱导的多种开放状态。”J.of Physiol.. 526. 47-56 (2000)

Kuzuya Masafumi: "VEGF Protects Toxicity of Oxidized LDL to Endothelial Cell via Intracellular Glutathione-Dependent Mechanism through KDR Receptor"Arteriosclerosis, Thrombosis, and Vascular Biology. (in press). (2001)

Kuzuya Masafumi：“VEGF 通过 KDR 受体通过细胞内谷胱甘肽依赖性机制保护氧化 LDL 对内皮细胞的毒性”动脉硬化、血栓形成和血管生物学。

Kuzuya Masafumi: "Glycation cross-links inhibit matrix metalloproteinase-2 activation in vascular smooth muscle cells cultured on collagen lattice."Diabetologia. (in press). (2001)

Kuzuya Masafumi：“糖化交联抑制在胶原晶格上培养的血管平滑肌细胞中基质金属蛋白酶-2 的活化。”Diabetologia。

Koike Teruhiko: "Activation of MMP-2 by Clostridium difficile Toxin B in bovine smooth muscle cells"Biochem.Biophys.Res.Commun. 277. 43-46 (2000)

Koike Teruhiko：“牛平滑肌细胞中艰难梭菌毒素 B 激活 MMP-2”Biochem.Biophys.Res.Commun。

Kuzuya M, Asai T, Kanda S, Maeda K, Cheng XW, Iguchi A.: "Glycation cross-links inhibit matrix metalloproteinase-2 activation in vascular smooth muscle cells cultured on collagen lattice."Diabetologia. (in press). (2001)

Kuzuya M、Asai T、Kanda S、Maeda K、Cheng XW、Iguchi A.：“糖化交联抑制胶原晶格上培养的血管平滑肌细胞中基质金属蛋白酶 2 的活化。”糖尿病学。