Spranger T, Finckh B, Fingerhut R, Kohlschütter A, Beisiegel U, Kontush A
Universitätskrankenhaus Eppendorf, Medizinische Kern und Poliklinik, Biochemisches Labor, Hamburg, Germany.
Chem Phys Lipids. 1998 Jan;91(1):39-52. doi: 10.1016/s0009-3084(97)00092-3.
Lipoprotein oxidation induced in vitro in whole plasma is expected to represent a more relevant model of the lipoprotein oxidation in the arterial wall than the in vitro oxidation of single isolated lipoproteins, e.g. low density lipoprotein (LDL). However, it remains unclear, how lipoprotein oxidation occurring in plasma is related to chemical composition and properties of the latter as well as to those of individual plasma lipoproteins. The present study was undertaken to characterize, how different constituents of human plasma contribute to the oxidizability of plasma lipoproteins oxidized directly in plasma samples. Oxidizability of plasma lipoproteins was assessed as oxidizability of whole heparin plasma and was measured spectrophotometrically as an increase in absorbance at 234 nm. To relate plasma oxidizability to its chemical composition and properties, plasma hydrophilic and lipophilic antioxidants, fatty acids, total lipids and TRAP were measured. To relate plasma oxidizability to the properties of individual lipoproteins, chemical composition and oxidizability were evaluated for LDL. We found that the oxidation kinetics of heparin plasma (diluted 150-fold and oxidized by 50 microM Cu2+) was characterized by three consecutive phases similar to the lag-, propagation and decomposition phases of LDL oxidation. Plasma oxidizability measured as different characteristics of these phases correlated negatively with plasma initial SH-groups, albumin, ascorbate, bilirubin, alpha-tocopherol, ubiquinol-10, free cholesterol, monounsaturated and saturated fatty acid content and positively with plasma initial total cholesterol, cholesterol ester and polyunsaturated fatty acid content. Plasma oxidizability measured as a rate of conjugated diene accumulation after different periods of oxidation correlated negatively with plasma initial albumin, urate, alpha-carotene and beta-carotene content. A positive correlation between oxidizabilities of whole plasma and LDL (isolated from the same plasma samples and oxidized by 14 mol Cu2+/mol LDL) was found. These data show that the oxidizability of plasma samples is critically determined by their chemical composition. They also suggest that the plasma oxidizability measured as an increase in absorbance at 234 nm may be used as a practical measure of the oxidizability of plasma lipoproteins.
与单个分离的脂蛋白(如低密度脂蛋白,LDL)的体外氧化相比,全血浆中体外诱导的脂蛋白氧化有望代表动脉壁中脂蛋白氧化的更相关模型。然而,血浆中发生的脂蛋白氧化如何与血浆的化学成分和特性以及单个血浆脂蛋白的化学成分和特性相关,仍不清楚。本研究旨在表征人血浆的不同成分如何影响直接在血浆样品中氧化的血浆脂蛋白的氧化能力。血浆脂蛋白的氧化能力通过全肝素化血浆的氧化能力来评估,并通过分光光度法测量234nm处吸光度的增加来测定。为了将血浆氧化能力与其化学成分和特性相关联,测量了血浆亲水和亲脂抗氧化剂、脂肪酸、总脂质和TRAP。为了将血浆氧化能力与单个脂蛋白的特性相关联,评估了LDL的化学成分和氧化能力。我们发现,肝素化血浆(稀释150倍并被50μM Cu2+氧化)的氧化动力学具有三个连续阶段,类似于LDL氧化的延迟、传播和分解阶段。作为这些阶段的不同特征测量的血浆氧化能力与血浆初始SH基团、白蛋白、抗坏血酸、胆红素、α-生育酚、泛醇-10、游离胆固醇、单不饱和脂肪酸和饱和脂肪酸含量呈负相关,与血浆初始总胆固醇、胆固醇酯和多不饱和脂肪酸含量呈正相关。作为不同氧化时间段后共轭二烯积累速率测量的血浆氧化能力与血浆初始白蛋白、尿酸盐、α-胡萝卜素和β-胡萝卜素含量呈负相关。发现全血浆和LDL(从相同血浆样品中分离并被14mol Cu2+/mol LDL氧化)的氧化能力之间存在正相关。这些数据表明,血浆样品的氧化能力关键取决于其化学成分。它们还表明,以234nm处吸光度增加测量的血浆氧化能力可作为血浆脂蛋白氧化能力的一种实用测量方法。