丁香乙酸乙酯相抑制LDL中赖氨酸、色氨酸氧化修饰的研究  

Inhibition effect of ethyl acetate fraction of clove on Trp and Lys modification during LDL oxidation

作　　者：万严 杨琼玉[1,2] 曲文娟[1,3] 谢丽琴[1] 郝澍[1,2] 沈勇根[1,4] 张良慧[1,2] 江慎华[1,2,3,4] 

机构地区：[1]九江学院药学与生命科学学院,江西九江332000 [2]九江安德和生物科技有限公司,江西九江332000 [3]江苏大学食品与生物工程学院,江苏省农产品物理加工重点实验室,江苏镇江212013 [4]江西农业大学食品科学与工程学院、江西省天然产物与功能食品重点实验室,江西南昌330045

出　　处：《食品工业科技》2017年第17期10-16,23,共8页Science and Technology of Food Industry

基　　金：国家自然科学基金(31360371,31560308,31301423);江西省科技支撑计划(20151BBF60026);江西省教育厅项目(GJJ161081);江苏省农产品物理加工重点实验室开放课题(JAPP2010-5);江西省天然产物与功能食品重点实验室开放基金;九江学院人才启动基金;九江学院教改课题(2015-04)

摘　　要：低密度脂蛋白(LDL)所含的色氨酸(Trp)、赖氨酸(Lys)发生氧化修饰是导致动脉粥样硬化(AS)形成的重要因素。为反映丁香乙酸乙酯相(EAFC)对LDL氧化过程中Trp、Lys修饰的抑制作用,本文首先对LDL氧化评价孵育体系中氧化剂Cu SO4及底物LDL浓度进行优化;进而以荧光为主要指标对这种抑制效果进行了评价。结果表明,选定CuSO_4浓度1.25、25μmol/L与LDL浓度500μg/m L为配比做后续实验。20μg/m L EAFC能抑制Trp降解导致的荧光淬灭,抑制作用显著高于1μg/m L阳性对照(2,6-二叔丁基对甲基苯酚,BHT)(p<0.01)。3μg/m L EAFC能抑制高反应活性醛修饰Lys,10μg/m L EAFC能显著抑制Lys与4-HNE共价结合(p<0.01)。15μg/m L EAFC能有效抑制Lys与MDA交联,抑制效果强于1μg/m L BHT。15μg/m L EAFC能显著抑制脂褐素形成(p<0.01)。本文为后期解析EAFC抑制LDL氧化修饰作用方式、抑制氧化LDL与清道夫受体之间识别等作用机理提供参考。Oxidative modification of Tryptophan （Trp） and Lysine （Lys） in apoB- 100 of LDL is the important factor resulting in atherosclerosis （AS）.In order to investigate the possible inhibition effect of Ethyl acetate fraction of clove （EAFC）on Trp and Lys oxidative modification during LDL oxidation, the concentrations of CuSO4 and LDL were firstly optimized, and then this inhibition effects of EAFC against oxidation were evaluated by using fluorescence as the main index in this paper.The results indicated that the concentrations of CuSO4 and LDL were 1.25,25 μmol/L and 500 μg/mL respectively in subsequent experiments.Fluorescence quenching can be inhibited by 20 μg/mL EAFC, and this inhibition effect was significantly higher than that of the 1 μg/mL butylated hydroxytoluene（ positive control ,BHT） （p 〈0.01 ）.Likewise ,3 μg/mL EAFC can inhibit the modification of Lys residues by highly reactive aldehydes.10 μg/mL EAFC can dramatically inhibit covalent binding of 4-HNE with the Lys residues（p 〈 0.01 ）.The modification of Lys residues resulting from MDA was inhibited by EAFC （15 μg/mL） which was significant than that of 1 μg/mL BHT.Meanwhile, 15 μg/mL EAFC showed a significant inhibition on formation of lipofuscin（p 〈0.01 ）.This study provides references for further analyzing the inhibition way of EAFC and the mechanism that EAFC inhibits the recognition between ox-LDL and scavenger receptors.

关 键 词：丁香 ApoB-100 低密度脂蛋白 色氨酸 赖氨酸 

分 类 号：TS201.4[轻工技术与工程—食品科学]