title: Dynamic aspects of ascorbic acid metabolism in the circulation: analysis by ascorbate oxidase with a prolonged in vivo half-life.
abstract: Because AA (L-ascorbic acid) scavenges various types of free radicals to form MDAA (monodehydroascorbic acid) and DAA (dehydroascorbic acid), its regeneration from the oxidized metabolites is critically important for humans and other animals that lack the ability to synthesize this antioxidant. To study the dynamic aspects of AA metabolism in the circulation, a long acting AOase (ascorbate oxidase) derivative was synthesized by covalently linking PEG [poly(ethylene glycol)] to the enzyme. Fairly low concentrations of the modified enzyme (PEG-AOase) rapidly decreased AA levels in isolated fresh plasma and blood samples with a concomitant increase in their levels of MDAA and DAA. In contrast, relatively high doses of PEG-AOase were required to decrease the circulating plasma AA levels of both normal rats and ODS (osteogenic disorder Shionogi) rats that lack the ability to synthesize AA. Administration of 50 units of PEG-AOase/kg of body weight rapidly decreased AA levels in plasma and the kidney without affecting the levels in other tissues, such as the liver, brain, lung, adrenal grand and skeletal muscles. PEG-AOase slightly, but significantly, decreased glutathione (GSH) levels in the liver without affecting those in other tissues. Suppression of hepatic synthesis of GSH by administration of BSO [L-buthionin-(S,R)-sulfoximine] enhanced the PEG-AOase-induced decrease in plasma AA levels. These and other results suggest that the circulating AA is reductively regenerated from MDAA extremely rapidly and that hepatic GSH plays important roles in the regeneration of this antioxidant.
authors.affiliation: Department of Biochemistry and Molecular Pathology, Osaka City University Medical School, 1-4-3 Asahimachi, Abeno, Osaka 545-8585, Japan. kasahara@med.osaka-cu.ac.jp
authors.forename: Emiko
authors.initials: E
authors.lastname: Kasahara
authors.forename: Misato
authors.initials: M
authors.lastname: Kashiba
authors.forename: Mika
authors.initials: M
authors.lastname: Jikumaru
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kuratsune
authors.forename: Kumi
authors.initials: K
authors.lastname: Orita
authors.forename: Yurika
authors.initials: Y
authors.lastname: Yamate
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Hara
authors.forename: Atsuo
authors.initials: A
authors.lastname: Sekiyama
authors.forename: Eisuke F
authors.initials: EF
authors.lastname: Sato
authors.forename: Masayasu
authors.initials: M
authors.lastname: Inoue
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001204:Ascorbate Oxidase / Q000737:chemistry / Q000378:metabolism*; D001205:Ascorbic Acid / Q000097:blood* / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D005978:Glutathione / Q000378:metabolism; D006207:Half-Life; D008099:Liver / Q000378:metabolism; D008297:Male; D010084:Oxidation-Reduction; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000975:Antioxidants; D011092:Polyethylene Glycols; D001204:Ascorbate Oxidase; D005978:Glutathione; D001205:Ascorbic Acid