【作者】 张波；

【导师】 李大鹏；

【作者基本信息】 山东农业大学 ， 食品科学， 2015， 硕士

【摘要】 苹果中的多酚类化合物品种繁多,含量丰富,在医药、食品等领域具有广阔的应用前景。本论文采用体外化学方法和细胞模型评价体系,研究了苹果中含有的几种多酚类化合物的抗氧化和抗炎症活性,并对其可能存在的交互作用进行了评价。主要研究内容和结果如下:1.采用氯化铝显色法测定了槲皮素、(+)-儿茶素、(-)-表儿茶素以及芦丁等4种黄酮类化合物单独使用及两两合用的总黄酮含量,并将合用后的测定值与理论值进行比较,建立评价交互作用的方法。实验结果表明,(+)-儿茶素与(-)-表儿茶素合用后,其总黄酮含量显著提高,说明该组合产生协同作用;(+)-儿茶素与芦丁的组合则与之相反,产生拮抗作用。其它组合测定值与理论值均未呈现显著性差异,表明这些组合在测定中呈现出加和效应。2.采用福林酚试剂法测定了槲皮素、(+)-儿茶素、(-)-表儿茶素、芦丁、5-咖啡酰奎宁酸、阿魏酸及没食子酸等7种多酚类化合物单独使用及两两合用的福林酚还原能力,并将测定值与理论值进行比较,建立评价交互作用的方法。实验结果表明,槲皮素与芦丁的组合、(-)-表儿茶素与5-咖啡酰奎宁酸的组合、芦丁与没食子酸的组合、5-咖啡酰奎宁酸与阿魏酸的组合,其福林酚还原能力实际值显著大于理论值,说明这些组合在测定时产生了协同作用;与之相反,(+)-儿茶素与(-)-表儿茶素的组合产生了拮抗作用。其他组合没食子酸当量浓度理论值和实际值无显著性差异,表明这些组合在测定中呈现加和作用。3.采用HepG2细胞模型研究了槲皮素、5-咖啡酰奎宁酸、(+)-儿茶素与(-)-表儿茶素的抗氧化活性。在分析单药抗氧化作用的基础上,采用1:1、1:3和3:1三种物质的量浓度比例进行两两组合实验,又将其中任意三种、四种化合物采用等物质的量浓度比例进行合用实验,研究这些组合可能存在的抗氧化交互作用。结果表明,多酚类化合物合用后,在部分比例浓度范围内产生协同抗氧化作用,随着组合中化合物种类的增加,产生协同作用的组合所占比例明显上升。4.采用脂多糖诱导小鼠巨噬细胞RAW264.7模型研究了槲皮素、5-咖啡酰奎宁酸、(+)-儿茶素与(-)-表儿茶素的抗炎症作用。在此基础上,又将其中任意两种、三种和四种成分按等物质的量浓度比例组合分析其抗炎症交互作用。结果表明,多酚类化合物合用后,在部分比例和浓度范围内产生对NO的协同抑制作用;随着组合中化合物种类的增加,产生协同作用的组合所占比例明显上升。更多还原

【Abstract】 Apple polyphenols are well known for their wide variety and abundance,and have broad application prospect in pharmaceutical,food and other fields.In this paper,in vitro chemical methods and cellular evaluation models were adopted to study the antioxidant and antiinflammation activities of several polyphenols in apple and their potential interactions.The main research contents and results are as follows:1.The total flavonoids contents of 4 kinds of flavonoids compounds,quercetin,(+)-catechin,(-)-epicatechin and rutin were determined by aluminum chloride method.Both individual compounds and their combinations were studied,and the measured values of the combinations were compared with the theoretical values.The results showed that when(+)-catechin and(-)-epicatechin were combined,their total flavonoids content increased significantly,indicating that synergistic effect appeared;on the contrary,in the combination of(+)-catechin and rutin,antagonistic effect was observed.There were no significant difference between the measured values and the theoretical values of other combinations,indicating that these combinations showed additive effect in the assay.2.Folin-Ciocalteu reducing capacities of 7 kinds of phenolic compounds,quercetin,(+)-catechin,(-)-epicatechin,rutin,5-caffeoylquinic acid and gallic acid were determined using the Folin-Ciocalteu reagent method.Both individual compounds and their combinations were studied,and the measured values of the combinations were compared with the theoretical values.The results showed that in the combinations of quercetin and rutin,(-)-epicatechin and 5-caffeoylquinic acid,rutin and gallic acid,and 5-caffeoylquinic acid and ferulic acid,the measured values of their Folin-Ciocalteu reducing capacity were significantly greater than the theoretical ones,indicating synergistic effect;on the contrary,when(+)-catechin and(-)-epicatechin were combined,antagonistic effect was observed.The measured values of other combinations showed no significant difference with the theoretical values,indicating that these combinations showed additive effect in the assay.3.In the Cellular Antioxidant Activity Assay,the antioxidant capacities of quercetin,5-caffeoylquinic acid,(+)-catechin and(-)-epicatechin were determined in a HepG2 cell model.After the determination of antioxidant effect of individual compounds,every two compounds in a molar concentration ratio of 1:1,1:3 and 3:1,and every three or four kinds of compounds in an equal molar concentration ratio were combined.The potential antioxidant interactions of these combinations were studied.The results showed that synergistic effect appeared in some combinations in certain molar proportions.With the increase in the compounds types in the combinations,more synergy combinations were observed.4.The anti-inflammation effect of quercetin,5-caffeoylquinic acid,(+)-catechin and(-)-epicatechin were determined in a lipopolysaccharide-induced mouse macrophage RAW264.7 model.On this basis,every two,three and four compounds in equal molar concentration were combined to study the potential anti-inflammatory interactions of these combinations.The results showed that,after the compounds were combined,synergistic NO inhibition effect appeared in some combinations in certain molar proportions.With the increase in the compounds types in combinations,more synergy combinations were observed.更多还原