The method for determination of serum lipid-free apolipoprotein concentration and its clinical application

血清无脂载脂蛋白浓度的测定方法及其临床应用

• 批准号: 11557204
• 负责人: SEISHIMA Mitsuru
• 金额: $ 6.91万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557204/
• 关键词: Lipid-free-free apolipoprotein; Corobary heart disease; Atherosclerosis; pre βHDL; Coronary heart disease; ELISA; preβ HDL

It is known that HDL are heterogeneous in particle size and small HDL particle can promote more effectively cholesterol efflux from cell membrane than large HDL particle. Small HDL are detected by two-dimentional gel electrophoresis, but this method is time consuming. Lipid-free apo A-I in this study was separated by HPLC and was elutred from 51 min to 56 min. The epitope of lipid-free apo A-I is the same as tjhat of a HDL, because we used the antibody to apo A-I in the ELISA system. We added SOU/ml of trasyrol to blood and also to 500 μ1 of fractionated solutions by HPLC. Serum lipid-free apo A-I concentration increased two times by repeated freeze and thawing test and changed with time even in the store at 4℃. Thus, serum sample was immediately treated for HPLC or freezed at -80℃ until use.Serum lipid-free apo A-I concentrations significantly increased in patients with coronary heart disease compared to healthy controls in both males and females. These findings show that reverse cholesterol transport is altered in coronary heat disease and the measurement of lipid-free apo A-I may be a good marker for atherosclerosis.

众所周知，HDL 的粒径是不均匀的，小 HDL 颗粒比大 HDL 颗粒更能有效地促进胆固醇从细胞膜流出。小HDL可以通过二维凝胶电泳来检测，但这种方法非常耗时。本研究中的无脂apo A-I通过HPLC分离，并在51分钟至56分钟洗脱。无脂apo A-I的表位与HDL的表位相同，因为我们在ELISA系统中使用了apo A-I的抗体。我们将 SOU/ml 的 trasyrol 添加到血液中，并通过 HPLC 添加到 500 μ1 分级溶液中。反复冻融试验，血清无脂apo A-I浓度增加两倍，即使在4℃保存，也随时间变化。因此，血清样品立即进行HPLC处理或-80℃冷冻备用。与健康对照相比，男性和女性冠心病患者血清中无脂apo A-I浓度均显着升高。这些发现表明，冠状动脉热病中胆固醇的反向转运发生了改变，无脂载脂蛋白 A-I 的测量可能是动脉粥样硬化的良好标志物。

项目成果

清島 満, 他: "高脂血症(上巻)リポ蛋白X"日本臨床.

Mitsuru Kiyoshima 等：“高脂血症（第 1 卷）脂蛋白 X”日本临床。

藤井秀比古 他: "医学のあゆみ「サイトカインと疾患」"医歯薬出版. 222 (2000)

Hidehiko Fujii 等：“医学史：细胞因子和疾病”Ishiyaku Publishing 222（2000）。

浦上克哉 他: "髄液中アポA-IおよびE"日本臨床. 57. 158-161 (1999)

Katsuya Urakami 等人：“脑脊液中的 ApoA-I 和 E”日本临床杂志 57. 158-161 (1999)。

Seishima,M., et al.: "Reference values of serum lipid"Rinsho Byori. 49. 1115-1121 (2001)

Seishima,M., et al.：“血清脂质参考值”Rinsho Byori。

Fei H, et al.: "Evaluation of two different homogeneous assays for LDL-cholesterol in lipoprotein-X-positive serum"Clin. Chem.. 46・9. 1351-1356 (2000)

Fei H 等人：“脂蛋白 X 阳性血清中 LDL-胆固醇的两种不同均相测定的评估”Clin. 1351-1356 (2000)。