LDL-Proteoglycans Complex as a fatty strek site in atherosclerosis.

低密度脂蛋白-蛋白聚糖复合物作为动脉粥样硬化中的脂肪运动部位。

• 批准号: 09670741
• 负责人: HAMADA Masanori
• 金额: $ 2.05万
• 依托单位: Wakayama Medical Collage
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670741/
• 关键词: 動脈硬化; glycosaminoglycans; low density lipoprotein

Objective : It has been reported that low density lipoprotein(LDL) produce insoluble complex with mono-sulfated glycosamino-glycans(GAGs). The complex also has a high affinity with calcium binding proteins namely osteopontin(OPN). We reported that the GAGs are increased in the aorta of experimental models of hypertension in rat. The main aim of this experiment was to investigate how the increased GAGs react with LDL to make insoluble GAGs-LDL complex at the site of atheroscleritic lesion and how the OPN is controled by angiotensin II.Design and Methods : 96-well microtiteation plate was coated with a certain amount of LDL. A limiting quantity of biotin-conjugated proteoglycans(PrG) is allowed to bind to each coated well in competition with different kind of GAGs or lipoproteins applied during incubation period. The amount of biotin-conjugated PrG retained on well was estimated spectrophotometrically by alkaline phosphatase-avidin method. The OPN produced in the culture medium by cultur … More ed vascular smooth muscle cells are measured by Western blotting with monoclonal antibody to OPN.Results : Lp(a) and very low density lipoprotein (VLDL) showed higher affinity with chondroitin-PrG than LDL and high density lipoprotein(HDL) but not significantly. Heparin showed significantly higher affinity with chondroitin-PrG than hiarulonic acid, heparan sulfate and chondroitin sulfate. Divalent cations(calcium, magnesium or manganese) were important to obtain insoluble GAGs-LDL complex in this in vitro model. Angiotensin II increased OPN protein in the medium in accordance with the concentrations between 10ィイD2-11ィエD2M to 10ィイD2-7ィエD2M. Whereas Sar-Ala angiotensin II did not. Angiotensin II receptor blockade supressed OPN increase produced by angiotensin II completely at a concentration of 10ィイD2-5ィエD2M.Conclusions : Our results showed that GAGs and divalent cations are essential for LDL to remain in the atherosclerotic lesion. Hypertension might contribute to make the complex by enhance production of chondroitin sulfate in the aortic media. Angiotensin II might enhance hydroxyapatite formation in hypertensive atherosclerotic lesion. Less

目的：据报道，低密度脂蛋白（LDL）与单硫酸化糖胺聚糖（GAG）产生不溶性复合物，该复合物还与钙结合蛋白即骨桥蛋白（OPN）具有高亲和力。大鼠高血压实验模型的主动脉中增加的GAGs如何与LDL反应以在该部位形成不溶性GAGs-LDL复合物。动脉粥样硬化病变以及血管紧张素 II 如何控制 OPN。 设计和方法：96 孔微滴板涂有一定量的 LDL，允许有限数量的生物素缀合蛋白聚糖 (PrG) 结合到每个涂层孔上。与孵育期间应用的不同种类的GAG或脂蛋白的竞争估计孔中保留的生物素缀合的PrG的量。采用分光光度法，使用 OPN 单克隆抗体，通过蛋白质印迹法测量培养血管平滑肌细胞在培养基中产生的 OPN。 结果：显示 Lp(a) 和极低密度脂蛋白 (VLDL)。与软骨素-PrG 的亲和力高于 LDL 和高密度脂蛋白 (HDL)，但与肝素的亲和力不显着较高。在该体外模型中，软骨素-PrG 比透明质酸、硫酸乙酰肝素和硫酸软骨素（钙、镁或锰）对于获得不溶性 GAG-LDL 复合物更重要，根据浓度，血管紧张素 II 会增加培养基中的 OPN 蛋白。 10 D2-11 D2M 至 10 D2-7 D2M 之间。 Sar-Ala 血管紧张素 II 在 10D2-5D2M 浓度下，血管紧张素 II 受体阻断完全抑制血管紧张素 II 产生的 OPN 增加。结论：我们的结果表明，GAG 和二价阳离子对于 LDL 留在动脉粥样硬化病变中至关重要。可能通过增强主动脉中层硫酸软骨素的产生来促进复合物的形成。 II 可能会增强高血压动脉粥样硬化病变中的羟基磷灰石形成。

项目成果

S.Booka,M.Hamada: "Glycosaminoglycans enhmces lipid accumulation in hypertension" J.of Hypertension. 61s. 321 (1998)

S.Booka，M.Hamada：“糖胺聚糖增强高血压中的脂质积累”J.of Hypertension。