基于Isobologram方法探究五倍子单宁与Vc、SOD的协同抗氧化作用

以五倍子单宁为主要原料,维生素C(Vc)、超氧化物歧化酶(SOD)为辅料,以1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除、2,2′-联氮-二(3-乙基苯并噻唑啉-6-磺酸)二铵盐(2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid,ABTS))自由基清除、铁离子还原能力(Ferricion Reducing Antioxidant Power,FRAP)、硫氰酸铁(Ferric Thocyanant,FTC)法等体外抗氧化试验进行比较,运用DPPH自由基清除试验结合等辐射分析法(Isobologram)探究五倍子单宁与Vc、SOD间的协同抗氧化作用。结果显示:在非油脂体系中,Isobologram分析图中五倍子单宁与Vc、五倍子单宁与SOD复配后的效应点均分布在相加线及95%可信限下方,各复配组相互作用指数(γ)均小于1,且实际IC_(50 mix)值均显著小于理论IC_(50 add)值,五倍子单宁与Vc、SOD间均存在协同效应,其中协同效果最强的复配组合比例分别为五倍子单宁:Vc=1∶3,五倍子单宁:SOD=1∶12。在油脂体系中,五倍子单宁、Vc和SOD的氧化抑制率分别为90.19±0.84%、11.84±1.15%、3.17±0.18%,五倍子单宁与Vc复配后不存在协同效应,氧化抑制率为60.23±1.55%;五倍子单宁与SOD复配后存在协同抗油脂氧化作用,氧化抑制率为94.58±2.21%,较五倍子单宁、SOD分别增加4.39%、91.41%。该试验结果为五倍子单宁系列协同抗氧化活性的研究以及开发复合抗氧化剂提供理论参考。

Isobologram Analysis of Synergistic Antioxidation of Chinese Gallotannins with Vc and SOD

In order to probe the synergistic antioxidant effects of Chinese Gallotannins with Vitamin C(Vc)or Superoxide Dismutase(SOD),Chinese Gallotannins was selected as raw material,Vc and SOD as auxiliary materials,and the antioxidant capacity of Chinese Gallotannins in the presence of Vc or SOD was evaluated by DPPH radical scavenging assay,ABTS radical scavenging assay,Ferricion Reducing Antioxidant Power,Ferric Thocyanant(FTC)method coupled with Isobologram analysis.The results showed that,in the non-oil system,the effect points of the combination of Chinese Gallotannins and Vc,and Chinese Gallotannins with SOD were all distributed below the additive line and 95% confidence limit in the Isobologram analysis diagram,the interaction index(γ)of these compound groups were less-than 1,and the actual IC_(50 mix) value was significantly less than the theoretical IC_(50 add) value.There was a synergistic effect between Chinese Gallotannins with Vc or SOD,The optimal combination proportion of synergetic effect were Chinese Gallotannins:Vc=1∶3 and Chinese Gallotannins:SOD=1∶12 respectively.In the oil system,the oxidation inhibition rates of Chinese Gallotannins,Vc and SOD were 90.19±0.84%,11.84±1.15% and 3.17±0.18% respectively.There was no synergistic effect between Chinese Gallotannins and Vc,and the oxidation inhibition rate was 60.23±1.55%.The combination of Chinese Gallotannins and SOD had synergistic anti-lipid oxidation effect,and the oxidation inhibition rate was 94.58±2.21%,which was 4.39% and 91.41% higher than that of Chinese Gallotannins and SOD,respectively.The results of this work provide fundamental experimental data reference to the development of synergistic antioxidant activities of Chinese Gallotannins.