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源自猪冠状动脉再生内皮的细胞含有更多氧化形式的载脂蛋白B-100,而氧化型低密度脂蛋白的摄取没有改变。

Cells derived from regenerated endothelium of the porcine coronary artery contain more oxidized forms of apolipoprotein-B-100 without a modification in the uptake of oxidized LDL.

作者信息

Kennedy Simon, Fournet-Bourguignon Marie-Pierre, Breugnot Christine, Castedo-Delrieu Maria, Lesage Ludovic, Reure Hélène, Briant Cyril, Leonce Stephane, Vilaine Jean-Paul, Vanhoutte Paul M

机构信息

Institut de Recherches Servier, Suresnes, France.

出版信息

J Vasc Res. 2003 Jul-Aug;40(4):389-98. doi: 10.1159/000072817. Epub 2003 Aug 6.

Abstract

Increased accumulation of lipoproteins and cholesterol within cells from regenerated endothelium may be responsible for their reported dysfunction. This study compared the presence and uptake of oxidized forms of low-density lipoprotein (LDL) in cells derived from native and regenerated endothelium. Four weeks after balloon denudation, primary cultures of native and regenerated endothelial cells were prepared from porcine coronary arteries. Regenerated endothelium stained more strongly using an antibody against oxidized lipoproteins. The increase in oxidized forms of apolipoprotein-B-100 exhibited by cells from regenerated endothelium was not due to an increase in extracellular-induced oxidation of native LDL, measured as the production of thiobarbituric-acid-reactive substances, being identical in both cell types. Intracellular cholesterol and cholesterol ester content were unchanged in regenerated cells. Using flow cytometry, accumulation of oxidized LDL was investigated further by quantifying the uptake of a mildly oxidized preparation of 1,1'-dioctadecyl-3,3,3',3-tetramethyl-indocarbocyanine perchlorate-labelled LDL. The parameters of uptake, EC(50) and E(max), were not different between cells from native and regenerated endothelium suggesting that the number of LOX-1 receptors was identical in the two cell types. Moreover, a negative correlation between the increased uptake of acetylated LDL and decreased cGMP production in response to bradykinin was observed in cells from regenerated endothelium. Thus, the increased incorporation of modified LDL and their intracellular oxidation could be responsible for the alteration in NO production. The presence of oxidized forms of LDL may be a marker of endothelium regeneration and could be involved in the endothelial dysfunction of pig coronary arteries 4 weeks after balloon denudation.

摘要

再生内皮细胞内脂蛋白和胆固醇积累的增加可能是其功能障碍的原因。本研究比较了天然内皮细胞和再生内皮细胞来源的细胞中氧化型低密度脂蛋白(LDL)的存在和摄取情况。球囊剥脱术后四周,从猪冠状动脉制备天然和再生内皮细胞的原代培养物。使用抗氧化脂蛋白抗体时,再生内皮细胞染色更强。再生内皮细胞来源的细胞中载脂蛋白B-100氧化形式的增加并非由于细胞外诱导的天然LDL氧化增加,两种细胞类型中硫代巴比妥酸反应性物质的产生相同,以此来衡量。再生细胞内胆固醇和胆固醇酯含量未发生变化。使用流式细胞术,通过定量摄取1,1'-二辛基-3,3,3',3-四甲基吲哚羰花青高氯酸盐标记的轻度氧化LDL制剂,进一步研究氧化LDL的积累情况。天然内皮细胞和再生内皮细胞来源的细胞之间摄取参数EC(50)和E(max)没有差异,这表明两种细胞类型中LOX-1受体的数量相同。此外,在再生内皮细胞来源的细胞中观察到,乙酰化LDL摄取增加与缓激肽刺激后cGMP产生减少之间存在负相关。因此,修饰LDL掺入增加及其细胞内氧化可能是一氧化氮产生改变的原因。LDL氧化形式的存在可能是内皮再生的标志物,并且可能参与球囊剥脱术后4周猪冠状动脉的内皮功能障碍。

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