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Vitamin C depletion increases superoxide generation in brains of SMP30/GNL knockout mice |
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| Authors: | Kondo Yoshitaka Sasaki Toru Sato Yasunori Amano Akiko Aizawa Shingo Iwama Mizuki Handa Setsuko Shimada Nobuko Fukuda Mitsugu Akita Masumi Lee Jaewon Jeong Kyu-Shik Maruyama Naoki Ishigami Akihito |
| Affiliation: | a Aging Regulation, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0015, Japan b Research Team for Molecular Biomarkers, Tokyo Metropolitan Institute of Gerontology, Tokyo 173-0015, Japan c Department of Biochemistry, Faculty of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba 274-8510, Japan d Division of Morphological Science, Biomedical Research Center, Saitama Medical School, Saitama 350-0495, Japan e Department of Pharmacy, College of Pharmacy, Longevity Life Science and Technology Institute, Pusan National University, Gumjeong-gu, Busan 609-735, Republic of Korea f College of Veterinary Medicine, Kyungpook National University, Daegu City 702-701, Republic of Korea |
| Abstract: | Vitamin C (VC) has a strong antioxidant function evident as its ability to scavenge superoxide radicals in vitro. We verified that this property actually exists in vivo by using a real-time imaging system in which Lucigenin is the chemiluminescent probe for detecting superoxide in senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice, which cannot synthesize VC in vivo. SMP30/GNL KO mice were given 1.5 g/L VC [VC(+)] for 2, 4, or 8 weeks or denied VC [VC(−)]. At 4 and 8 weeks, VC levels in brains from VC(−) KO mice were <6% of that in VC(+) KO mice. Accordingly, superoxide-dependent chemiluminescence levels determined by ischemia-reperfusion at the 4- and 8 weeks test intervals were 3.0-fold and 2.1-fold higher, respectively, in VC(−) KO mice than in VC(+) KO mice. However, total superoxide dismutase activity and protein levels were not altered. Thus, VC depletion specifically increased superoxide generation in a model of the living brain. |
| Keywords: | EDTA, ethylenediaminetetraacetic acid GNL, gluconolactonase HPLC, high-performance liquid chromatography KO, knockout ODS, osteogenic disorder Shionogi ROS, reactive oxygen species SDS, sodium dodecyl sulfate SMP30, senescence marker protein-30 SOD, superoxide dismutase VC, vitamin C WT, wild type |
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