| Co-Investigator(Kenkyū-buntansha) |
NAGAOKA Shin-ichi Ehime University, Faculty of Science, Professor, 理工学研究科, 教授 (30164403)
OHARA Keishi Ehime University, Faculty of Science, PhD., 理工学研究科, 助手 (10284390)
TAJIMA Kunihiko Kyoto Institute of Technology, Department of Applied Biology, Professor, 繊維学部, 教授 (50163457)
MITO Masaki Kyusyu Institute of Technology, Faculty of Engineering, Associate Professor, 工学部, 助教授 (60315108)
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| Budget Amount *help |
¥40,800,000 (Direct Cost: ¥40,800,000)
Fiscal Year 2006: ¥10,200,000 (Direct Cost: ¥10,200,000)
Fiscal Year 2005: ¥8,500,000 (Direct Cost: ¥8,500,000)
Fiscal Year 2004: ¥10,200,000 (Direct Cost: ¥10,200,000)
Fiscal Year 2003: ¥11,900,000 (Direct Cost: ¥11,900,000)
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| Research Abstract |
Human aging is induced by the reaction of lipid, protein, and nucleic acid with active oxygens and free radicals. Natural antioxidants such as vitamin E (α-, β-, γ-, δ-tocopherol, TocH), ubiquinol, vitamin C, polyphenols (flavones, catechins) etc., which exist in biological systems, scavenge active oxygens and free radicals quickly, and protect the aging. In the present work, the kinetic studies of i) free-radical-scavenging action, ii) tocopherol-regeneration action, and iii) singlet-oxygen-quenching action with the above natural antioxidants were performed, and several interesting results were obtained.
1) The aroxyl-radical-scavenging rates (k_s) of vitamin E (α-, β-, γ-, δ-TocH) and ubiquinol were measured in micellar solutions by varying pH, using stopped-flow spectrophotometer. It has been clarified that ubiquinol shows higher and lower free-radical-scavenging activity than a-tocopherol in mitochondria of several tissues (heart, muscle, liver and brain) and in serum, respectively.
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2) Quenching rates (K_Q) of singlet oxygen (^1O_2) by 7 kinds of flavone derivatives and 4 kinds of catechins have been measured The K_Q values of several kinds of flavones and catechins were found to be larger than those of lipids, amino acids, and DNA. Further, these values are similar to those of a-and y-tocopherol, ubiquinol-10, and plastoquinol model, suggesting that these polyphenols may contribute to the protection of oxidative damage in biological systems, by quenching ^1O_2.
3) In order to understand the dynamics of antioxidant actions of vitamin E (α-,β-, γ-, δ-TocH) in biological systems, kinetic study of the formation and decay reactions of vitamin E radicals (α-, β-, γ-, δ-tocopheroxyls, Toc) has been performed in organic solvents, using stopped-flow spectrophotometer. The second-order rate constants (k_f and 2k_d) for the formation and decay reactions of Toc were determined by comparing the observed curves with the simulation ones obtained by the numerical calculation of differential equations related to the above reactions. The k_f values decreased in the order of α-> β-~γ-> δ-TocH, and the 2kd values increased in the order of α-< β-< γ-< δ-Toc in ethanol. The wavelengths of absorption maxima (λmax^1) and molar extinction coefficients (ε_1) of the optical spectra (for example, i = 1~4 for α-Toc.) were determined for these radicals. The scheme of the decay reactions of α-, β-, γ-, δ-Toc radicals has been discussed based on the results obtained.
4)The rate constant and activation energy of the regeneration reaction of natural vitamin E by vitamin C were determined with a double-mixing
stopped-flow spectrophotometer. The kinetic effect ofmethyl substitution on the aromatic ring of vitamin E radical indicates that partial charge-transfer plays a role in the reaction Since a substantial deuterium kinetic isotope effect was not found, the tunneling effect may not play an important role under the present experimental conditions. Less
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