Application of enzymatic fluorometric assays to quantify phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in human plasma lipoproteins
Application of enzymatic fluorometric assays to quantify phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in human plasma lipoproteins
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DOI：
10.1016/j.chemphyslip.2021.105102
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发表时间：
2021-06-09
影响因子：
3.4
通讯作者：
Morita, Shin-ya
中科院分区：
文献类型：
--
作者：
Tsuji, Tokuji;Yuri, Tatsushi;Morita, Shin-ya
磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）和鞘磷脂（SM）是血浆脂蛋白的重要表面成分，包括极低密度脂蛋白（VLDL）、低密度脂蛋白（LDL）和高密度脂蛋白（HDL）。然而，由于量化脂蛋白中磷脂类别的困难，PC、PE和SM在脂蛋白中的病理生理学作用尚未得到很好的表征。在这项研究中，我们评估了酶荧光测定法测量 VLDL、LDL 和 HDL 中 PC、PE 和 SM 的精密度和准确度，这些物质是通过超速离心从人血浆中分离出来的。脂蛋白中 PC、PE 和 SM 测量的批内变异系数 (CV) 分别为 1.5-2.8%、1.1-2.4% 和 0.9-2.3%，而 PC、PE 和 SM 测定的批间 CV 分别为 2.7-4.7%、2.1-4.5% 和 1.6-3.3%。测量 VLDL、LDL 和 HDL 中 PC、PE 和 SM 的所有测定均实现了出色的线性和几乎完全回收。我们使用这些酶荧光测定的初步结果表明，VLDL、LDL 和 HDL 的磷脂组成不同。总之，我们建立了高通量酶荧光测定法来定量人血浆 VLDL、LDL 和 HDL 中的 PC、PE 和 SM，这将有助于进一步研究磷脂在脂蛋白中的病理生理作用。
Phosphatidylcholine (PC), phosphatidylethanolamine (PE) and sphingomyelin (SM) are important surface components of plasma lipoproteins, including very-low-density lipoproteins (VLDL), low-density lipoproteins (LDL) and high-density lipoproteins (HDL). However, the pathophysiological roles of PC, PE and SM in lipoproteins have not been well characterized owing to the difficulties in quantifying phospholipid classes in lipoproteins. In this study, we assessed the precision and accuracy of the enzymatic fluorometric assays for measuring PC, PE and SM in VLDL, LDL and HDL, which were isolated from human plasma by ultracentrifugation. The within-run coefficients of variation (CV) for the measurements of PC, PE and SM in lipoproteins were 1.5-2.8 %, 1.1-2.4 % and 0.9-2.3 %, respectively, whereas the between-run CVs for the PC, PE and SM assays were 2.7-4.7 %, 2.1-4.5 % and 1.6-3.3 %, respectively. Excellent linearity and almost complete recovery were achieved for all assays measuring PC, PE and SM in VLDL, LDL and HDL. Our preliminary results using these enzymatic fluorometric assays suggested that the phospholipid compositions were different among VLDL, LDL and HDL. In conclusion, we established high-throughput enzymatic fluorometric assays to quantify PC, PE and SM in human plasma VLDL, LDL and HDL, which will be useful for further investigation of pathophysiological roles of phospholipids in lipoproteins.