Arginyl-fructose (AF) and arginyl-fructosyl-glucose (AFG) were chemically synthesized and purified. Their in vitro and cellular antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity assay, respectively. The peroxyl radical scavenging ...
Arginyl-fructose (AF) and arginyl-fructosyl-glucose (AFG) were chemically synthesized and purified. Their in vitro and cellular antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity assay, respectively. The peroxyl radical scavenging activity of AF was much higher than that of AFG, which was in good agreement with their reduction capacity to donate electrons or hydrogen atoms. On the other hand, the hydroxyl radical scavenging activity of AF was weaker than that of AFG, which was consistent with their metal chelating activity, suggesting that AFG-$Cu^{2+}$ complex may be less redox-active than AF-$Cu^{2+}$ complex due to 1 glucose molecule attached. The cellular antioxidant activity of AF and AFG appeared to depend on both their permeability into cell membrane and the scavenging activity on peroxyl or hydroxyl radicals. These results indicate that AF and AFG, Maillard reaction products, may have a high potential as a material for the development of nutraceutical food with antioxidant activity.
Arginyl-fructose (AF) and arginyl-fructosyl-glucose (AFG) were chemically synthesized and purified. Their in vitro and cellular antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity assay, respectively. The peroxyl radical scavenging activity of AF was much higher than that of AFG, which was in good agreement with their reduction capacity to donate electrons or hydrogen atoms. On the other hand, the hydroxyl radical scavenging activity of AF was weaker than that of AFG, which was consistent with their metal chelating activity, suggesting that AFG-$Cu^{2+}$ complex may be less redox-active than AF-$Cu^{2+}$ complex due to 1 glucose molecule attached. The cellular antioxidant activity of AF and AFG appeared to depend on both their permeability into cell membrane and the scavenging activity on peroxyl or hydroxyl radicals. These results indicate that AF and AFG, Maillard reaction products, may have a high potential as a material for the development of nutraceutical food with antioxidant activity.
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가설 설정
In this study, we prepared AF and AFG by chemical synthesis, investigated the in vitro and cellular antioxidant activity of AF and AFG, and discussed what kind of the mechanism might be involved in their in vitro and cellular antioxidant activity.
제안 방법
The ORAC assay was carried out on a Tecan GENios multifunctional plate reader (Salzburg, Austria) with fluorescent filters (excitation wavelength: 485 nm and emission filter:535 nm). In the final assay mixture, fluorescein (40 nM) was used as a target of free radical attack with either AAPH (20 mM) as a peroxyl radical generator in peroxyl radical scavenging capacity (ORACROO·) assay (11) or H2O2-CuSO4 (H2O2, 0.
To determine whether the peroxyl radical scavenging· activity of them could result from their reduction capacity donating electrons or hydrogen to peroxyl radical, the ability of AF and AFG to stimulate the reduction of copper ion was investigated by measuring the concentration of Cu+ formed from Cu2+ through the reduction by electrons.
Louis, MO, USA). HepG2 cell was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).
데이터처리
Statistical analyses were done using statistical package SPSS (Statistical Package for Social Science, SPSS Inc., Chicago, IL, USA) program, and significance of each group was verified with the analysis of one-way analysis of variance (ANOVA) followed by Student’s t-test for comparison of means.
The significance of differences between sample and AAPH treated group at p<0.05*, 0.01**, and 0.001*** was determined with Student’s t-test.
The significance of differences between sample and Cu2+ treated group at **p<0.01 was determined with Student’s t-test.
이론/모형
Determination of reduction capacity The reducing capacity of AF and AFG was determined according to the method of Aruoma et al. (13). The 40 µL of different concentrations of AF or AFG in distilled water were mixed with 160 µL of the mixture containing 0.
Intracellular ROS measurement using DCFH-DA assay Cellular oxidative stress owing to ROS generating by AAPH or Cu2+ was measured by spectrofluorometrically by the DCFH-DA method (15). DCFH-DA diffuses through the cell membrane and is enzymatically hydrolyzed by intracellular esterase to the non-fluorescent DCFH, which can be rapidly oxidized to the highly fluorescent DCF in the presence of ROS.
성능/효과
In conclusion, the peroxyl radical scavenging activity of AF was much higher than that of AFG, which was in good agreement with their reduction pacacity to donate electrons or hydrogen atoms. On the other hand, the hydroxyl radical scavenging activity of AF was weaker than that of AFG, which was consistent with their metal chelating activity, suggesting that AFG-Cu2+ complex may be less redoxactive than AFi-Cu2+ complex due to 1 glucose molecule attached.
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