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Functional characterization of HDL in patients with type 2 diabetes using simple separation method of plasma HDL fraction

使用血浆 HDL 组分的简单分离方法表征 2 型糖尿病患者的 HDL 功能

基本信息

批准号：
22790519
负责人：
TANABE Jutaro
金额：
$ 1.08万
依托单位：
Hirosaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2010
资助国家：
日本
起止时间：
2010 至 2011
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22790519/
关键词：
臨床検査医学代謝学内分泌学

项目摘要

We developed a simple method to separate HDL fraction from human plasma, by which several properties of HDL function were possible to be evaluated simultaneously from plasma in type 2 diabetic patients.Subjects were 14 patients with type 2 diabetes mellitus (9 males, 5 females, age 59.3 ±9.8 y.o.) who were admitted to our department. Laboratory data were HbA1c 8.4±1.2%, TC 181.6±26.2mg/dl, LDL-C 110.4±25.7 mg/dl, HDL-C 41.9±8.6mg/dl and TG 114.4±54.4mg/dl. Fasting plasma sample was applied to polyacrylamide gel electrophoresis (PAGE); Lipophor, Jokoh, Japan, then sharp HDL fraction was recovered using Phoreto-yield protein collection device (Jokoh, Japan) equipped with cut-off MW 15,000. In this procedure, native HDL fraction was obtained within 1 hour. We analyzed several functional markers as follows in native HDL fraction simultaneously :(1) carboxymethyl-lysin (CML) for glucoxidation, (2) paraoxonase-1 (PON-1) activity for anti-oxidation (3) serum amyroid A protein (SAA) for inflmmation.We evaluated the accuracy of each analytical method. The values in HDL fraction were compensated with those in plasma levels. Recovery rate of HDL from plasma was corrected with ApoA-I conc. and about 70%. CV (coefficient of variation) % in HDL recovery, PON-1 activity and SAA in plasma and HDL fraction were about 25 %.1) CML level in HDL fraction was not detected different from plasma (0.0574±0.0331 mg/mL).2) PON-1 activities in HDL fraction (19.5±5.78 x104 U/L/min)were higher than those in plasma (4.14±2.36 x104 U/L/min).3) SAA levels in HDL fraction (15.8±31.6 ng/mL) were the same as those in plasma (18.1±21.6 ng/mL).These findings demonstrated that the ability of HDL to inhibit glycation and lipid oxidation may reflect changes in specific enzyme and protein components in HDL particles.

我们建立了一种从血浆中分离高密度脂蛋白组分的简便方法，可同时测定2型糖尿病患者血浆中高密度脂蛋白（HDL）的多种功能。被我们部门录取了实验室数据为HbA 1c 8.4± 1.2%，TC 181.6±26.2mg/dl，LDL-C 110.4± 25.7mg/dl，HDL-C 41.9±8.6mg/dl和TG 114.4±54.4mg/dl。将空腹血浆样品应用于聚丙烯酰胺凝胶电泳（PAGE）; Lipophor，Jokoh，Japan，然后使用配备有截留分子量15，000的Phoreto-产量蛋白收集装置（Jokoh，Japan）回收尖锐的HDL级分。在该程序中，在1小时内获得天然HDL级分。我们同时分析了天然HDL组分中的几种功能标志物：（1）用于葡萄糖氧化的羧甲基溶素（CML），（2）用于抗氧化的对氧磷酶-1（PON-1）活性，（3）用于炎症的血清淀粉样蛋白A（SAA），并对每种分析方法的准确性进行了评价。HDL分数的值用血浆水平的值补偿。血浆HDL回收率用ApoA-I浓度校正，约为70%。CV HDL回收率的（变异系数）%，血浆和HDL组分中的PON-1活性和SAA约为25%。1）HDL组分中CML水平与血浆无明显差异（0.0574±0.0331 mg/mL）。2）HDL组分中PON-1活性（19.5±5.78 × 104 U/L/min）高于血浆（4.14±2.36 × 104 U/L/min）。3）HDL组分中的SAA水平（15.8±31.6 ng/mL）与血浆中的浓度相同这些结果表明HDL抑制糖基化和脂质氧化的能力可能反映了HDL颗粒中特定酶和蛋白质组分的变化。