摘    要：本研究旨在探讨葡萄籽提取物对高级脂肪氧化终产物（Advanced lipid peroxidation end products，ALEs）的抑制效果。首先构建添加葡萄籽提取物（Grape Seed Extract，GSE）的羟基自由基催化氧化猪肉肌原纤维蛋白（porcine myofibrillar protein，MP）的食品体系，然后通过测定体系中次级氧化产物（MDA)的含量、脂褐质类荧光物质含量、蛋白质的羰基含量、巯基含量、表面疏水性、内源色氨酸荧光强度、体外消化率等，研究GSE对体系中ALEs的抑制效果。本研究发现GSE的添加使得脂肪氧化程度、脂褐质类荧光物质含量、蛋白羰基含量和MDA相关的ALEs含量等随着GSE浓度的增大而逐渐降低；同时MHC进一步降解;GSE使得MP表面疏水性有所回升，而巯基含量则在氧化的基础上进一步降低;色氨酸内源荧光进一步逐渐降低;MP的体外消化率在添加GSE后进一步降低。

Abstract：This study was to investigate the inhibitory effect of grape seed extract on Advanced lipid peroxidation end products (ALEs). Firstly, a food system for the oxidation of porcine myofibrillar protein (MP) catalyzed by hydroxyl radicals was prepared by Grape Seed Extract (GSE), The GSE system was then studied by measuring the content of secondary oxidation products (MDA), lipofuscin phosphors, protein carbonyl content, thiol content, surface hydrophobicity, endogenous tryptophan fluorescence intensity, and in vitro digestibility to evaluate the inhibitory effect of ALEs. This study found that the addition of GSE caused the degree of lipid oxidation, lipofuscin-like fluorescent substance content, protein carbonyl content and MDA-related ALEs content to gradually decrease with the increase of GSE concentration; meanwhile, MHC further degraded; GSE made MP surface hydrophobic Sex increased, but the thiol content further decreased on the basis of oxidation; the endogenous fluorescence of tryptophan decreased further; the in vitro digestibility of MP decreased further after GSE addition.

关键词：葡萄籽提取物； 猪肉肌原纤维蛋白； 氧化； 高级脂肪氧化终产物

Keywords: grape seed extract; porcine myofibrillar protein; oxidation; advanced lipid peroxidation end product 

目    录

1 引言 1

2 材料与仪器 2

2.1 实验材料与试剂 2

2.2 实验仪器与设备 3

3 实验方法 3

3.1 猪肉肌原纤维蛋白的提取 3

3.2建立MP氧化生成ALEs体系 4

3.3脂质氧化的测定 4

3.4体系中脂褐质类荧光物质的测定 4

3.5蛋白羰基含量的测定 4

3.6 SDS-PAGE分析 4

3.7蛋白巯基含量的测定 5

3.8蛋白表面疏水性的测定 5

3.9蛋白内源色氨酸荧光光谱测定 5

3.10数据统计和分析 6

4 结果分析 6

4.1不同浓度的GSE对体系中脂质氧化的影响 6

4.2不同浓度的GSE处理对脂褐质类荧光物质的影响 6

4.3不同浓度的GSE处理对氧化后MP羰基含量的影响 7

4.4氧化与 GSE添加对蛋白交联的影响 8

4.5不同浓度的GSE处理对氧化后MP巯基含量的影响