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肝素化前血浆中脂蛋白和肝脂酶活性的测定及其生理意义

Measurement and physiological significance of lipoprotein and hepatic lipase activities in preheparin plasma.

作者信息

Watson T D, Tan C E, McConnell M, Clegg S K, Squires L F, Packard C J

机构信息

Department of Pathological Biochemistry, Glasgow Royal Infirmary, University NHS Trust, UK.

出版信息

Clin Chem. 1995 Mar;41(3):405-12.

PMID:7882516
Abstract

A radiochemical method for selective measurement of postheparin lipase activities was adapted to analyze lipoprotein lipase and hepatic lipase in preheparin plasma. The assay sensitivity was increased about four-fold by doubling both the volume of plasma used and the volume of lipolytic products taken for liquid scintillation counting, and was further improved by increasing the incubation period by 50% to 90 min. Rabbit antiserum to human hepatic lipase was unsuitable for the selective measurement of lipoprotein lipase because of apparent endogenous lipolytic activity. Preheparin hepatic lipase, however, was sensitive to inactivation by sodium dodecyl sulfate (SDS), the inhibition being greatest (> 90%) for plasma incubated with an equal volume of 40 mmol/L SDS. Intra- and interassay CVs for the two enzymes were 12.5-14.6% and 17.4-19.7%, respectively. In a cross-sectional study of 84 healthy subjects, pre- and postheparin hepatic lipase activities were higher in men than women, were correlated with indices of obesity, and were significantly correlated with one another, which explained the association of the former with plasma concentrations of high-density lipoprotein (HDL), HDL2, and small, dense low-density lipoproteins. There was no significant relationship between pre- and postheparin lipoprotein lipase activities, but the former were correlated with plasma concentrations of free fatty acids (FFA) and very-low-density lipoprotein. Apparently, preheparin activities of hepatic lipase, but not of lipoprotein lipase, may be a useful measure of the physiological function of "whole body" enzyme activity in cross-sectional and metabolic studies, where heparinization is not possible. Preheparin lipoprotein lipase activities, however, may reflect displacement of the enzyme by FFA and subsequent binding to remnants of triglyceride-rich lipoproteins.

摘要

一种用于选择性测量肝素后脂肪酶活性的放射化学方法被应用于分析肝素前血浆中的脂蛋白脂肪酶和肝脂肪酶。通过将所用血浆体积和用于液体闪烁计数的脂解产物体积都加倍,测定灵敏度提高了约四倍,并且通过将孵育时间延长50%至90分钟进一步得到改善。兔抗人肝脂肪酶血清由于明显的内源性脂解活性,不适用于脂蛋白脂肪酶的选择性测量。然而,肝素前肝脂肪酶对十二烷基硫酸钠(SDS)的失活敏感,对于与等体积40 mmol/L SDS孵育的血浆,抑制作用最大(>90%)。两种酶的批内和批间变异系数分别为12.5 - 14.6%和17.4 - 19.7%。在一项对84名健康受试者的横断面研究中,肝素前和肝素后肝脂肪酶活性男性高于女性,与肥胖指标相关,且彼此显著相关,这解释了前者与高密度脂蛋白(HDL)、HDL2以及小而密低密度脂蛋白血浆浓度的关联。肝素前和肝素后脂蛋白脂肪酶活性之间无显著关系,但前者与游离脂肪酸(FFA)和极低密度脂蛋白的血浆浓度相关。显然,在无法进行肝素化的横断面和代谢研究中,肝素前肝脂肪酶活性而非脂蛋白脂肪酶活性可能是“全身”酶活性生理功能的有用测量指标。然而,肝素前脂蛋白脂肪酶活性可能反映了该酶被FFA取代并随后与富含甘油三酯脂蛋白残粒结合的情况。

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