异鼠李素及橙皮甙抑制HDL氧化修饰作用的研究

Inhibitory Effect of Isorhamnetin and Hesperidin on the Oxidation of High-density Lipoproteins (HDL) Induced by Cu^(2+)

目的研究异鼠李素及橙皮甙对高密度脂蛋白(HDL)氧化修饰的抑制作用。方法按一次性密度梯度超速离心法分离人血清HDL,分别加入5.0μmol/L异鼠李素及橙皮甙温育3h,然后利用Cu2+介导体外氧化修饰脂蛋白的方法,观察HDL氧化修饰不同时间,其234nm吸光度(A234)、相对电泳迁移率(REM)、硫代巴比妥酸反应物质(TBARS)及蛋白质羰基的动力学改变。结果Cu2+介导HDL氧化修饰2、4、6、8、12及24h后,其A234、REM、TBARS及蛋白质羰基含量均逐步增加;加入5.0μmol/L异鼠李素及橙皮甙的实验组,Cu2+介导HDL的氧化修饰分别延迟2~6h和2~4h;其A234、REM、TBARS及载脂蛋白羰基含量较对照组降低,异鼠李素实验组分别降低16.3%~46.9%、0.6%~25.2%、9.2%~28.4%及11.6%~45.2%;橙皮甙实验组降低1.5%~30.0%、1.4%~13.3%、6.6%~18.8%及14.4%~62.0%。结论异鼠李素及橙皮甙可抑制Cu2+诱导的HDL的氧化修饰,且异鼠李素的抗氧化作用比橙皮甙强。

Objective To test the inhibitory effect of isorhamnetin and hesperidin on the HDL oxidation induced by Cu^2＋. Methods The serum HDL was isolated by the one step density gradient ultracentrifugation. The HDL oxidation was induced by Cu^2＋ in vitro for different length of periods. Isorhamnetin and hesperidin at 5 μmol/L, respectively, were added 3 h before the copper oxidation. The oxidation of HDL was measured by the levels of absorbance at 234 nm （A234）, relative electrophoretic mobility （REM）, thiobarbituric acid reactive substances （TBARS） and protein carbonyls content. Results （1）A234, REM, TBARS and protein carbonyls formation increased gradually after 2, 4, 6, 8, 12 and 24 h of HDL oxidation induced by Cu^2＋ without treatment with isorhamnetin or hesperidin. （2）With the treatment of isorhamnetin or hesperidin, the kinetic changes of A234, REM, TBARS and protein carbonyls formation delayed 2-6 h and 2-4 h, respectively. Compared with the controls, the A234, REM, TBARS and protein carbonyls formation were reduced by 16.3%-46. 9% （P〈0. 001）, 0.6%-25.2% （P〈0.05）, 9. 2%-28.4% （P〈0.01） and 11.6%-45.2% （P〈0.001） by isorhamnetin and by 1.5%-30. 0%, 1. 4%-13. 3%, 6. 6%-18. 8% （P〈0. 05） and 14. 4%-62. 0% （P〈0. 001） by hesperidin, respectively. Conclusion Both isorhamnetin and hesperidin inhibit the oxidation of HDL, but isorhamnetin functions stronger.