| Project/Area Number |
17590064
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | Prefectural University of Hiroshima |
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Principal Investigator |
MIWA Nobuhiko Prefectural University of Hiroshima, Faculty of Life and Environmental Sciences, Professor (00142141)
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| Co-Investigator(Kenkyū-buntansha) |
SAITOH Yasukazu Prefectural University of Hiroshima, Faculty of Life and Environmental Sciences, Associate Lecturer (90405514)
XIAO Li 県立広島大学, 生命環境学部 三羽研究室, ラボ主任研究員 (10405516)
河地 信哉 県立広島大学, 生命環境学部 三羽研究室, ラボ主任研究員 (70438290)
GAN Xuehui 県立広島大学, 大学院・総合学術研究科, 研究員 (20405517)
谷 孝二 県立広島大学, 三羽研究室, 研究員 (30405518)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,710,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Vitamin C regeneration / dhar Gene / Reactive oxygen species / Hydroperoxide / Cell death defense / Hydrogen-dissolved water / dhar遺伝子 / 細胞死防御 / デヒドロアスコルビン酸 / グルタチオン / 低酸素-再酸素化 / 細胞死 / DNA切断 / 酸化ストレス軽減 / 細胞死制御 / ビタミンC細胞内取込み / 虚血-再灌流障害 |
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| Research Abstract |
1. In Chinese hamster ovary cells CHO that were transfected with a dehydroascorbic acid (DehAsc) reductase (dhar) gene-incorporated pRcCMV vector, DHAR proteins were expressed scarcely in the nuclei and distributed all over the cytoplasm where DHAR was absent for dhar-nontransfected cells, and the peroxylipid-induced cell mortality and DNA strand cleavages were demonstrated to be prevented by simultaneous administration with DehAsc and the glutathione derivative GSH-iPr. 2. In the post-ischemic reperfusion model, hypoxia and subsequent reoxygenation (H/R) system, cell mortality was markedly occurred for dhar-nontransfected cells, and prevented for dhar-transfected cells by DehAsc and GSH-iPr which were simultaneously administered at 2-5 hr before hypoxia treatment. 3. Simultaneous DehAsc/ GSH-iPr administration to CHO/dhar cells achieved, at 1-6 hr after the administration, enrichment of intracellular Asc, which markedly repressed intracellular hydroperoxides and hydrogen peroxides tha
… More
t increased either after the peroxylipid t-BuOOH treatment or drastically at 1-3 min after post-hypoxic reoxygenation with DehAsc/GSH-iPr. 4. Why have a dhar gene been evolutionarily conserved whereas the Asc-synthesis rate-determining enzyme gene GLO was omitted at a stage of the primates during biological evolution? In hydrogen-dissolved water with reducing properties such as a dissolved-hydrogen concentration of 963 ppb and a redox potential of -788 mV, Asc-recycling and intracellular uptake of DehAsc were not promoted in correspondence to diminished ROS (reactive oxygen species) amounts in CHO/dhar cells, and intracellular ROS was appreciably alleviated, in contrast to abundant ROS for hydrogen-deficient pure water-prepared culture media. Residual rates of Asc at 27 hr after aqueous dissolution were diminished to 20-30% versus the initial concentration for pure water, and retained over 70% for hydrogen-dissolved water. Thus a reductive environment like hydrogen-dissolved water decreased a need for dhar gene expression resulting in Asc regeneration, and consequently achieved so-called "gene saving" as purposiveness. In summary, evolutionary conservation of dhar gene in contrast to omission of GLO gene might be attributed to no realization for retention of reducing conditions such as hydrogen-dissolved water for 3.8 billion years of biology evolution, and efficient Asc-regeneration and ROS-repression were not satisfied by endogenous dhar gene, and necessitated transfer of exogenous dhar gene as shown in the present study. Less
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