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浙江大学学报（医学版）
浙江大学学报（医学版）  2018, Vol. 47 Issue (1): 27-34    DOI: 10.3785/j.issn.1008-9292.2018.02.04
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绿原酸抑制低密度脂蛋白非酶糖基化和氧化修饰研究
蔡蕊1(),陈枢青1,*(),江慎华2,*()
1. 浙江大学药学院, 浙江 杭州 310058
2. 九江学院药学与生命科学学院, 江西 九江 332005
Chlorogenic acid inhibits non-enzymatic glycation and oxidation of low density lipoprotein
CAI Rui1(),CHEN Shuqing1,*(),JIANG Shenhua2,*()
1. College of Pharmacy, Zhejiang University, Hangzhou 310058, China
2. School of Pharmacy and Life Science, Jiujiang University, Jiujiang 332005, Jiangxi Province, China
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摘要：

目的: 研究绿原酸对人体低密度脂蛋白（LDL）非酶糖基化和氧化修饰的抑制作用。方法: 建立LDL非酶糖基化孵育体系，采用紫外-可见分光光度法测定糖基化早期产物（Amodori产物）和中期产物（二羰基化合物）的含量，荧光分光光度计测定糖基化末期产物的含量；建立LDL氧化孵育体系，采用紫外-可见分光光度法测定硫代巴比妥酸反应物（TBARS）和共轭二烯的含量，荧光分光光度法测定色氨酸荧光淬灭强度以及脂褐素、总荧光产物、活性醛和丙二醛的含量，并进一步通过三维荧光等高线特征谱验证。结果: 在LDL糖基化修饰模型中，150 μg/mL和300 μg/mL的绿原酸均能够抑制Amodori产物、二羰基化合物和糖基化末期产物的生成；在LDL氧化修饰模型中，15 μg/mL和25 μg/mL的绿原酸均能够抑制TBARS的生成；5 μg/mL和10 μg/mL的绿原酸对色氨酸荧光淬灭，以及对活性醛、丙二醛、总荧光产物、脂褐素和共轭二烯的生成均有抑制作用。三维荧光等高线特征谱结果与前一致。结论: 绿原酸能够抑制LDL非酶糖基化和氧化修饰。
关键词： 绿原酸/药理学;  脂蛋白类, LDL/药物作用;  糖基化/药物作用;  光谱分析;  紫外线;  光谱法, 荧光/方法;  抗氧化剂    
Abstract:

Objective: To investigate the effect of chlorogenic acid (CGA) on non-enzymatic glycation and oxidation of low density lipoprotein (LDL). Methods: The non-enzymatic glycation incubation system of LDL-glucose was established. The contents of early glycation products (Amodori product) and intermediate products (dicarbonyl compound) were determined by ultraviolet-visible spectrophotometry, and the content of advanced glycation end products (AGEs) was determined by fluorescence spectrophotometry. The LDL oxidation incubation system was established. The contents of thiobarbituric acid reactive substances(TBARS) and conjugated diene were determined by ultraviolet-visible spectrophotometry. The tryptophan fluorescence quenching, and the content of lipofuscin, total fluorescence products, active aldehydes and malondialdehyde were determined by fluorescence spectrophotometry, and further verified by three-dimensional fluorescence spectroscopy. Results: In the LDL glycation experiment, 150 μg/mL and 300 μg/mL CGA inhibited the formation of Amadori product, dicarbonyl compounds and AGEs. In the LDL oxidation experiment, 15 μg/mL and 25 μg/mL CGA inhibited the formation of TBARS effectively; 5 μg/mL and 10 μg/mL CGA inhibited tryptophan fluorescence quenching, and the formation of active aldehydes, malondialdehyde, total fluorescence products, lipofuscin and conjugated diolefine. And the three-dimensional fluorescence spectroscopy showed the same results. Conclusion: CGA can inhibit non-enzymatic glycation and oxidation of LDL.
Key words: Chlorogenic acid/pharmacology    Lipoproteins, LDL/drug effects    Glycosylation/drug effects    Spectrum analysis    Ultraviolet rays    Spectrometry, fluorescence/methods    Antioxidants
收稿日期: 2017-12-30 出版日期: 2018-06-12
CLC:  R96  
基金资助: 国家自然科学基金(31360371);江西省重点研发计划(20171BBF60049);九江学院教学改革研究课题(2015-04);九江学院人才启动基金;九江市杰出青年人才计划
通讯作者: 陈枢青,江慎华     E-mail: cairuiangle@gmail.com;chenshuqing@zju.edu.cn;jiangshenhua66@163.com
作者简介: 蔡蕊(1992-), 女, 硕士研究生, 主要从事微生物与生化药学研究; E-mail:cairuiangle@gmail.com; https://orcid.org/0000-0002-1761-1209
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蔡蕊
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引用本文:

蔡蕊,陈枢青,江慎华. 绿原酸抑制低密度脂蛋白非酶糖基化和氧化修饰研究[J]. 浙江大学学报（医学版）, 2018, 47(1): 27-34.

CAI Rui,CHEN Shuqing,JIANG Shenhua. Chlorogenic acid inhibits non-enzymatic glycation and oxidation of low density lipoprotein. J Zhejiang Univ (Med Sci), 2018, 47(1): 27-34.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2018.02.04    或    http://www.zjujournals.com/med/CN/Y2018/V47/I1/27

图 1  绿原酸抑制低密度脂蛋白氧化修饰过程中活性醛和丙二醛的生成
图 2  三维荧光等高线特征谱表征绿原酸对低密度脂蛋白氧化修饰的抑制效果
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