Nourooz-Zadeh J, Tajaddini-Sarmadi J, Ling K L, Wolff S P
Department of Medicine, University College London Medical School, U.K.
Biochem J. 1996 Feb 1;313 ( Pt 3)(Pt 3):781-6. doi: 10.1042/bj3130781.
High-density lipoprotein (HDL) has been proposed as the principal carrier of hydroperoxides in plasma, based upon data gathered with an HPLC-chemiluminescence technique. To test this hypothesis we have measured total lipid hydroperoxides in native plasma using the ferrous oxidation in Xylenol Orange (FOX) assay and then fractionated plasma into very-low-density lipoprotein (LDL) and HDL fractions. Hydroperoxides were found to accumulate principally (more than 65%) in LDL, as judged by hydroperoxide content per amount of protein or cholesterol, or expressed as a proportion of total hydroperoxide in plasma. Plasma was also incubated at 37 degrees C in the presence and absence of 2,2'-azo-bis-(2-amidinopropane) hydrochloride (AAPH), an azo-initiator of lipid peroxidation. The majority of hydroperoxides generated in plasma were recovered in the LDL fraction. Furthermore, when isolated lipoproteins were subject to oxidation initiated by AAPH, very-low-density lipoprotein and LDL showed the greatest propensity for hydroperoxide accumulation, whereas HDL seemed relatively resistant. Estimates for plasma and LDL peroxidation based upon techniques which measure total lipid hydroperoxides suggest that levels of hydroperoxides in plasma and LDL are far higher than that those estimates generated by ostensibly more selective techniques. Higher levels of hydroperoxides in LDL than those reported by HPLC-chemiluminescence also seem in greater accordance with other available data concerning LDL oxidation.
基于用高效液相色谱-化学发光技术收集的数据,高密度脂蛋白(HDL)被认为是血浆中氢过氧化物的主要载体。为了验证这一假设,我们使用二甲酚橙亚铁氧化法(FOX法)测量了天然血浆中的总脂质氢过氧化物,然后将血浆分离为极低密度脂蛋白(VLDL)和高密度脂蛋白部分。根据每单位蛋白质或胆固醇中的氢过氧化物含量,或以血浆中总氢过氧化物的比例来判断,发现氢过氧化物主要(超过65%)积聚在VLDL中。血浆还在有无脂质过氧化偶氮引发剂2,2'-偶氮二(2-脒基丙烷)盐酸盐(AAPH)的情况下于37℃孵育。血浆中产生的大部分氢过氧化物在VLDL部分中被回收。此外,当分离的脂蛋白受到AAPH引发的氧化作用时,极低密度脂蛋白和低密度脂蛋白显示出最大的氢过氧化物积聚倾向,而高密度脂蛋白似乎相对具有抗性。基于测量总脂质氢过氧化物的技术对血浆和低密度脂蛋白过氧化的估计表明,血浆和低密度脂蛋白中的氢过氧化物水平远高于表面上更具选择性的技术所产生的估计值。低密度脂蛋白中氢过氧化物的水平高于高效液相色谱-化学发光法所报告的水平,这似乎也与其他有关低密度脂蛋白氧化的现有数据更为一致。