{"title":"暴露于铜的人血清中脂蛋白二烯形成的分析。","authors":"J Regnström, K Ström, P Moldeus, J Nilsson","doi":"10.3109/10715769309056514","DOIUrl":null,"url":null,"abstract":"<p><p>The susceptibility of low density lipoprotein (LDL) to oxidative modification can be determined by analyzing the lag phase for initiation of diene formation in isolated LDL exposed to Cu2+. However, the applicability of this assay for clinical studies is limited by the requirement of a preparative ultracentrifugation of LDL and that the influence of water soluble antioxidants and other lipoproteins is not accounted for. The present paper describes a modification of this assay allowing determination of lag phase for lipoprotein diene formation in serum. The formation of dienes in serum exposed to Cu2+ begins following the consumption of serum alpha-tocopherol, correlates to the formation of thiobarbituric acid reactive substances (r = 0.987, n = 8), is inhibited by the addition of ascorbic acid and is absent in lipoprotein-deficient serum. It is also accompanied by an increased mobility of serum lipoproteins on agarose gel electrophoresis and with an ability of serum to displace isolated copper-oxidized LDL from binding sites mediating degradation in mouse peritoneal macrophages. The coefficient of variance of the analysis is below 3%. It is concluded that this technique allows analysis of lipoprotein oxidation susceptibility in serum samples and may prove to be useful in clinical analysis of the lipoprotein oxidation susceptibility.</p>","PeriodicalId":12438,"journal":{"name":"Free radical research communications","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10715769309056514","citationCount":"85","resultStr":"{\"title\":\"Analysis of lipoprotein diene formation in human serum exposed to copper.\",\"authors\":\"J Regnström, K Ström, P Moldeus, J Nilsson\",\"doi\":\"10.3109/10715769309056514\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The susceptibility of low density lipoprotein (LDL) to oxidative modification can be determined by analyzing the lag phase for initiation of diene formation in isolated LDL exposed to Cu2+. However, the applicability of this assay for clinical studies is limited by the requirement of a preparative ultracentrifugation of LDL and that the influence of water soluble antioxidants and other lipoproteins is not accounted for. The present paper describes a modification of this assay allowing determination of lag phase for lipoprotein diene formation in serum. The formation of dienes in serum exposed to Cu2+ begins following the consumption of serum alpha-tocopherol, correlates to the formation of thiobarbituric acid reactive substances (r = 0.987, n = 8), is inhibited by the addition of ascorbic acid and is absent in lipoprotein-deficient serum. It is also accompanied by an increased mobility of serum lipoproteins on agarose gel electrophoresis and with an ability of serum to displace isolated copper-oxidized LDL from binding sites mediating degradation in mouse peritoneal macrophages. The coefficient of variance of the analysis is below 3%. It is concluded that this technique allows analysis of lipoprotein oxidation susceptibility in serum samples and may prove to be useful in clinical analysis of the lipoprotein oxidation susceptibility.</p>\",\"PeriodicalId\":12438,\"journal\":{\"name\":\"Free radical research communications\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1993-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.3109/10715769309056514\",\"citationCount\":\"85\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Free radical research communications\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.3109/10715769309056514\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Free radical research communications","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3109/10715769309056514","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 85
摘要
低密度脂蛋白(LDL)对氧化修饰的易感性可以通过分析暴露于Cu2+的低密度脂蛋白启动二烯形成的滞后期来确定。然而,这种检测方法在临床研究中的适用性受到低密度脂蛋白制备超离心要求的限制,而且水溶性抗氧化剂和其他脂蛋白的影响也没有考虑在内。本文描述了一种改进的方法,允许测定血清中脂蛋白二烯形成的滞后期。暴露于Cu2+的血清中,二烯的形成始于血清α -生育酚的消耗,与硫代巴比妥酸反应物质的形成相关(r = 0.987, n = 8),被添加抗坏血酸抑制,在脂蛋白缺乏的血清中不存在。它还伴随着琼脂糖凝胶电泳上血清脂蛋白流动性的增加,以及血清从小鼠腹膜巨噬细胞中介导降解的结合位点取代分离的铜氧化LDL的能力。分析的方差系数在3%以下。结果表明,该技术可用于血清样品中脂蛋白氧化敏感性的分析,并可用于脂蛋白氧化敏感性的临床分析。
Analysis of lipoprotein diene formation in human serum exposed to copper.
The susceptibility of low density lipoprotein (LDL) to oxidative modification can be determined by analyzing the lag phase for initiation of diene formation in isolated LDL exposed to Cu2+. However, the applicability of this assay for clinical studies is limited by the requirement of a preparative ultracentrifugation of LDL and that the influence of water soluble antioxidants and other lipoproteins is not accounted for. The present paper describes a modification of this assay allowing determination of lag phase for lipoprotein diene formation in serum. The formation of dienes in serum exposed to Cu2+ begins following the consumption of serum alpha-tocopherol, correlates to the formation of thiobarbituric acid reactive substances (r = 0.987, n = 8), is inhibited by the addition of ascorbic acid and is absent in lipoprotein-deficient serum. It is also accompanied by an increased mobility of serum lipoproteins on agarose gel electrophoresis and with an ability of serum to displace isolated copper-oxidized LDL from binding sites mediating degradation in mouse peritoneal macrophages. The coefficient of variance of the analysis is below 3%. It is concluded that this technique allows analysis of lipoprotein oxidation susceptibility in serum samples and may prove to be useful in clinical analysis of the lipoprotein oxidation susceptibility.