氧化、丙二醛修饰低密度脂蛋白的ELISA检测法及临床应用

Development of ELISA for oxidized,malondialdehyde-modified LDL and clinical value

目的建立血浆氧化、修饰低密度脂蛋白(LDL)ELISA检测法并进行临床研究。方法采用自制的多克隆抗体建立ELISA分别测定铜离子氧化型(Ox)和丙二醛(MDA)修饰型LDL,并对方法进行考核;对冠心病(CHD)患者、键康对照人群Ox-LDL、MDA-LDL水平进行分析。结果抗Ox-LDL与MDA-LDL几乎没有反应性;Ox-LDL和MDA-LDL测定平均批内变异(CV)分别为6.1%和6.8%,平均批间CV分别为9.7%和10.1%;Ox-LDL和MDA-LDL测定平均回收率分别为95.1%和93.7%;两法检测限均为:0.05～1.5mg/L。CHD患者较正常人血浆Ox-LDL[(178.1±73.8)mg/Lvs.(82.7±29.1)mg/L,P<0.01]、MDA-LDL[(48.7±25.6)mg/Lvs.(39.2±32.9)mg/L,P<0.05]水平均升高;Ox-LDL作为预测动脉粥样硬化发生指标的价值优于MDA-LDL。Ox-LDL、MDA-LDL分别同TC、TG、LDL-C呈正相关,而与HDL-C呈负相关。结论建立的方法特异、灵敏、准确,适于临床检测。高Ox-LDL、MDA-LDL与CHD密切相关。

Objective To develop enzyme-linked immunosorbent assays （ELISAs） for copper-oxidized （Ox）, malondialdehyde-modified （MDA） low-density lipoprotein （LDL） and study their clinical value. Methods Two ELISAs were developed for detecting Ox-LDL and MDA-LDL levels by using self-made polyclonal antibodies, whose precision, recovery and specificity were also tested. Ox-LDL and MDA-LDL levels were determined in patients with coronary heart disease （CHD） and healthy control subjects. Results Both Ox-LDL （ 178. 1 ± 73.8 mg/L vs. 82.7 ± 29. i mg/L,P 〈0.01 ） and MDA-LDL （48.7 ＋ 25.6 mg/L.vs. 39.2 ± 32.9 mg/L,P 〈 0.05） levels were increased in patients with CHD （ P 〈 0.05 ）. Ox-LDL and MDA-LDL levels were positively related with total cholesterol, triglyceride and LDL cholesterol, while negatively related with high-density lipoprotein. Conclusions The developed ELISAs are specific, pre- cise, accurate, and useful for clinical application. Both Ox-LDL and MDA-LDL are strong risk factors for atherosclerosis. As a risk factor for predicting atherosclerosis, Ox-LDL is better than MDA-LDL in this study.