STRUCTURAL-CHANGES OF HIGH-DENSITY-LIPOPROTEIN APOLIPOPROTEINS FOLLOWING INCUBATION WITH HUMAN POLYMORPHONUCLEAR CELLS
STRUCTURAL-CHANGES OF HIGH-DENSITY-LIPOPROTEIN APOLIPOPROTEINS FOLLOWING INCUBATION WITH HUMAN POLYMORPHONUCLEAR CELLS
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DOI：
10.1111/j.1432-1033.1994.tb18947.x
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发表时间：
1994-06-15
期刊：
影响因子：
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通讯作者：
MOATTI, N
中科院分区：
文献类型：
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作者：
COGNY, A;PAUL, JL;MOATTI, N
基于动脉粥样硬化的发病机制和事件与炎症反应的相似性，我们研究了人多形核白细胞(PMN)脱颗粒和氧化过程对高密度脂蛋白(高密度脂蛋白)结构的影响。在有或没有刺激剂的情况下，将高密度脂蛋白与中性粒细胞按生理比例(370nmol胆固醇-高密度脂蛋白/ml和2×10(6)PMN/ml)孵育15、30和60min。经SDS/PAGE和电聚焦后免疫印迹分析载脂蛋白(APE)发现：(A)载脂蛋白AII和载脂蛋白Cs的缓慢水解性；(B)载脂蛋白E的快速水解性；(C)载脂蛋白AI异构体分布的改变，最酸性的异构体(AI-2)增加，以较弱的酸性形式(AI-1)为代价；(D)主要载脂蛋白AII亚型转变为两种更基本的形式。相比之下，没有注意到硫代巴比妥酸反应物质(TBARS)评估的可量化的脂质修饰或脂质氧化。尽管TBARS没有变化，但观察到高密度脂蛋白维生素E含量下降了80%。由于在培养液中加入超氧化物歧化酶阻止了这种下降，我们得出结论，影响HDL的氧化过程发生了。用蛋白水解剂实验表明，弹性蛋白酶引起载脂蛋白E、AII和Cs的蛋白水解性裂解。相反，apo AI修饰可能既涉及氧化过程，也涉及蛋白质降解过程。
Based on the analogy in mechanisms and events between the pathogenesis of atherosclerosis and the inflammatory reaction, we investigated the impact of human polymorphonuclear leukocyte (PMN) degranulation and oxidative process on high-density-lipoprotein (HDL) structure.HDL were incubated (37 degrees C) with PMN at a physiological ratio (370 nmol cholesterol-HDL/ml with 2x10(6) PMN/ml) for 15, 30 and 60 min with or without stimulating agent. PMN activation was assessed by measurement of superoxide anion generation and elastase production, which both reached peak concentration at 15 min.HDL apolipoproteins (ape) analysed by immunoblotting after SDS/PAGE and electrofocusing evidenced the following modifications: (a) a slow hydrolysis of apo AII and apo Cs; (b) a rapid hydrolysis of apo E; (c) a change in apo AI isoform distribution with an increase in the most acidic isoform (AI-2) at the expense of a less acidic form (AI-1); (d) a shift of the major apo AII isoform into two more basic forms.In contrast, no quantifiable lipid modification nor lipid oxidation, assessed by thiobarbituric-acid-reactive substances (TBARS) were noted. Despite a lack of variation of TBARS, a decrease in HDL vitamin E content by 80% was observed. Since this decrease was prevented by addition of superoxide dismutase in the medium, we concluded the occurence of an oxidative process affecting HDL.Experiments with proteolytic inhibitors showed that elastase caused the proteolytic cleavage of apolipoprotein E, AII and Cs. In contrast, apo AI modification might involve both oxidative and proteolytic processes.