Harada Amane, Toh Ryuji, Murakami Katsuhiro, Kiriyama Maria, Yoshikawa Keiko, Miwa Keiko, Kubo Takuya, Irino Yasuhiro, Mori Kenta, Tanaka Nobuaki, Nishimura Kunihiro, Ishida Tatsuro, Hirata Ken-Ichi
Central Research Laboratories, Sysmex Corporation, Kobe, Japan.
Division of Evidence-Based Laboratory Medicine and.
J Appl Lab Med. 2017 Sep 1;2(2):186-200. doi: 10.1373/jalm.2016.022913.
Recent studies have shown that the cholesterol efflux capacity of HDL is a better predictor of cardiovascular disease (CVD) than HDL cholesterol. However, the standard procedures used for measuring cholesterol efflux capacity involve radioisotope-labeled cholesterol and cultured macrophages. Thus, a simpler method to measure HDL functionality is needed for clinical application.
We established a cell-free assay system to evaluate the capacity of HDL to accept additional cholesterol, which we named cholesterol "uptake capacity," using fluorescently labeled cholesterol and an anti-apolipoprotein A1 antibody. We quantified cholesterol uptake capacity of apolipoprotein B (apoB)-depleted serum samples from patients with coronary artery disease who had previously undergone revascularization.
This assay system exhibited high reproducibility (CV <10%) and a short processing time (<6 h). The myeloperoxidase-mediated oxidation of apoB-depleted serum impaired cholesterol uptake capacity. Cholesterol uptake capacity correlated significantly with cholesterol efflux capacity (r2 = 0.47, n = 30). Furthermore, cholesterol uptake capacity correlated inversely with the requirement for revascularization because of recurrence of coronary lesions in patients with optimal control of LDL cholesterol (P < 0.01, n = 156). A multivariate analysis adjusted for traditional coronary risk factors showed that only cholesterol uptake capacity remained significant (odds ratio, 0.48; 95% CI, 0.29-0.80; P = 0.0048).
Cholesterol uptake capacity assay evaluates the functionality of HDL in a sensitive and high-throughput manner without using radioisotope label and cells. This assay system could be used for the assessment of CVD risk in the clinical settings.
近期研究表明，高密度脂蛋白（HDL）的胆固醇流出能力比HDL胆固醇更能预测心血管疾病（CVD）。然而，用于测量胆固醇流出能力的标准程序涉及放射性同位素标记的胆固醇和培养的巨噬细胞。因此，临床应用需要一种更简单的方法来测量HDL功能。
我们建立了一种无细胞分析系统，使用荧光标记的胆固醇和抗载脂蛋白A1抗体来评估HDL接受额外胆固醇的能力，我们将其命名为胆固醇“摄取能力”。我们对先前接受过血运重建的冠心病患者的载脂蛋白B（apoB）缺乏血清样本的胆固醇摄取能力进行了量化。
该分析系统具有高重现性（CV<10%）且处理时间短（<6小时）。髓过氧化物酶介导的apoB缺乏血清氧化损害了胆固醇摄取能力。胆固醇摄取能力与胆固醇流出能力显著相关（r2 = 0.47，n = 30）。此外，在低密度脂蛋白胆固醇得到最佳控制的患者中，胆固醇摄取能力与因冠状动脉病变复发而进行血运重建的需求呈负相关（P < 0.01，n = 156）。对传统冠心病危险因素进行校正的多变量分析显示，只有胆固醇摄取能力仍然具有显著性（优势比，0.48；95%CI，0.29 - 0.80；P = 0.0048）。
胆固醇摄取能力分析以灵敏且高通量的方式评估HDL功能，无需使用放射性同位素标记和细胞。该分析系统可用于临床环境中CVD风险的评估。
