Abstract
The reduction of 20 μM hexavalent chromium [chromium(VI)] byl-ascorbic acid (AsA) (0.06–2 mM) and/or glutathione (GSH) (2–15 mM) in buffer solutions, cell-free bronchoalveolar lavage fluids or soluble fractions of rat lungs was investigated at physiological pH (37° C). The reduction in AsA solution was pseudo-first-order in a single phase with respect to chromium(VI), but that in GSH solution showed a two-phase process. The half-life of chromium(IV) ranged from seconds to hours. The reducing ability of AsA was markedly higher than that of GSH. Coexistence of equimolar GSH with AsA accelerated the reduction rate slightly, in comparison with that in the corresponding AsA solution. Lavage fluids containing 0.06 mM AsA showed pH-dependent reactions similar to those of the corresponding AsA solutions. The lungsoluble fractions reduced chromium(VI) in a process composed of phase I and phase II, characterized by the reducing ability of AsA-GSH cooperation and of AsA alone, respectively. Reduction in the former was 30–40% more rapid than in the latter. The biological half-life of chromium(VI) in the lung was estimated to be 0.6 min, on the basis of the reducing activity in the first phase. However, the apparent biological half-life of chromium(VI) was about 2 min in rat lungs after intratracheal injection of chromate, involving depletion of AsA, but no significant changes in GSH. The difference is discussed in terms of AsA-induced initiative reduction in the alveolar lining fluid and subsequent obstructive effects of the resulting trivalent species on trans-membrane permeability of chromate anions. These results suggest that AsA is more reactive than GSH in the reduction of chromium(VI) in the rat lung and that the extracellular AsA in the alveolar lining fluid plays an important role in antioxidant defense against inhaled chromium(VI) compounds.
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References
Adachi S, Yoshimura H, Miyayama R, Katayama H, Takemoto K, Kawai H (1983) Effects of chromium compounds on the respiratory system. Part 2. Difference between water-soluble hexavalent compounds and trivalent compounds. Jpn J Ind Health 25: 149–154
Baldea I, Munteanu L (1980) The kinetics of the oxidation of ascorbic acid by chromate. Studia Univ Babes-Bolyai Chemia 25: 24–31
Campomar JA, Fiol JJ, Terron A, Moreno V (1986) Chromium(III) interactions with nucleotides. II. Inorg Chim Acta 124: 75–81
Cantin AM, North SL, Hubbard RC, Crystal RG (1987) Normal alveolar epithelial lining fluid contains high levels of glutathione. J Appl Physiol 63: 152–157
Chatterjee IB (1973) Evolution and the biosynthesis of ascorbic acid. Science 182: 1271–1272
Clark JH (1959) The denaturation of proteins by chromium salts. A M A Arch Ind Health 20: 117–123
Connett PH, Wetterhahn KE (1983) Metabolism of the carcinogen chromate by cellular constituents. In: Structure and bonding. Inorganic elements in biochemistry. Springer-Verlag, Berlin, 54: 93–124
Connett PH, Wetterhahn KE (1985) In vitro reaction of the carcinogen chromate with cellular thiols and carboxylic acids. J Am Chem Soc 107: 4282–4288
DePamphilis ML, Cleland WW (1973) Preparation and properties of chromium(III)-nucleotide complexes for use in the study of enzyme mechanisms. Biochemistry 12: 3714–3724
Franchini I, Magnani F, Mutti A (1983) Mortality experience among chromeplating workers. Scand J Work Environ Health 9: 247–252
Garcia JD, Jennette KW (1981) Electron-transport cytochrome P-450 system is involved in the microsomal metabolism of the carcinogen chromate. J Inorg Biochem 14: 281–295
Ginter E (1980) Endogenous ascorbic acid synthesis and recommended dietary allowances for vitamin C. Am J Clin Nutr 33: 1448–1449
Gray SJ, Sterling K (1950) The tagging of red cells and plasma proteins with radioactive chromium. J Clin Invest 29: 1604–1613
Gruber JE, Jennette KW (1978) Metabolism of the carcinogen chromate by rat liver microsomes. Biochem Biophys Res Commun 82: 700–706
Hornig D (1975) Distribution of ascorbic acid, metabolites and analogues in man and animals. In: King CG, Burns JJ (ed) Second conference on vitamin C. Ann NY Acad Sci 258: 103–118
Keeling PL, Smith LL (1982) Relevance of NADPH depletion and mixed disulphide formation in rat lung to the mechanism of cell damage following paraquat administration. Biochem Pharmacol 31: 3243–3249
Keller DA, Menzel DB (1985) Picomole analysis of glutathione, glutathione disulfide, glutathione S-sulfonate, and cysteine S-sulfonate by high-performance liquid chromatography. Anal Biochem 151: 418–423
Kitagawa S, Seki H, Kametani F, Sakurai H (1982) Uptake of hexavalent chromium by bovine erythrocytes and its interaction with cytoplasmic components; the role of glutathione. Chem Biol Interact 40: 265–274
Korallus U, Harzdorf C, Lewalter J (1984) Experimental bases for ascorbic acid therapy of poisoning by hexavalent chromium compounds. Int Arch Occup Environ Health 53: 247–256
Legg JI (1978) Substitution-inert metal ions as probes of biological function. Coordination Chem Rev 25: 103–132
Léonard A, Lauwerys RR (1980) Carcinogenicity and mutagenicity of chromium. Mutat Res 76: 227–239
Levis AG, Bianchi V (1982) Mutagenic and cytogenetic effects of chromium compounds. In: Langård S (ed) Biological and environmental aspects of chromium. Elsevier Biomedical Press, Amsterdam, New York, Oxford, pp 171–208
Levis AG, Bianchi V, Tamino G, Pegoraro B (1978) Cytotoxic effects of hexavalent and trivalent chromium on mammalian cells in vitro. Br J Cancer 37: 386–396
Mikalsen A, Alexander J, Ryberg D (1989) Microsomal metabolism of hexavalent chromium. Inhibitory effect of oxygen and involvement of cytochrome P-450. Chem Biol Interact 69: 175–192
Nielson DW, Goerke J, Clements JA (1981) Alveolar subphase pH in the lungs of anesthetized rabbits. Proc Natl Acad Sci USA 78: 7119–7123
Norseth T (1981) The carcinogenicity of chromium. Environ Health Perspect 40: 121–130
Rucker RB, Dubick MA, Mouritsen J (1980) Hypothetical calculations of ascorbic acid synthesis based on estimates in vitro. Am J Clin Nutr 33: 961–964
Samitz MH (1970) Ascorbic acid in the prevention and treatment of toxic effects from chromates. Acta Derm Venereol (Stockholm) 50: 59–64
Scarpelli EM (1977) The surfactant system of the lung. Int Anesthesiol Clin 15: 19–60
Stubbs DW, Griffin JF (1973) The influence of dietary protein on gulonolactone hydrolase, gulonate NADP oxidoreductase, and tissue ascorbate in male and female rats. Proc Soc Exp Biol Med 144: 199–202
Suzuki Y (1988) Reduction of hexavalent chromium by ascorbic acid in rat lung lavage fluid. Arch Toxicol 62: 116–122
Suzuki Y, Fukuda K (1989) Anion-exchange high-performance liquid Chromatographic determination of ascorbic acid and hexavalent chromium in rat lung preparations after treatment with sodium chromate in vitro and in vivo. J Chromatogr 489: 283–290
Suzuki Y, Homma K, Minami M, Yoshikawa H (1984) Distribution of chromium in rats exposed to hexavalent chromium and trivalent chromium aerosols. Ind Health 22: 261–277
Tandon SK (1982) Organ toxicity of chromium in animals. In: Langård S (ed) Biological and environmental aspects of chromium. Elsevier Biomedical Press, Amsterdam, New York, Oxford, pp 209–220
Wiegand HJ, Ottenwälder H, Bolt HM (1984 a) The reduction of chromium(VI) to chromium(III) by glutathione: an intracellular redox pathway in the metabolism of the carcinogen chromate. Toxicology 33: 341–348
Wiegand HJ, Ottenwälder H, Bolt HM (1984 b) Disposition of intratracheally administered chromium(III) and chromium(VI) in rabbits. Toxicol Lett 22: 273–276
Willis RJ, Kratzing CC (1974) Ascorbic acid in rat lung. Biochem Biophys Res Commun 59: 1250–1253
Willis RJ, Kratzing CC (1976) Extracellular ascorbic acid in lung. Biochim Biophys Acta 444: 108–117
Yassi A, Nieboer E (1988) Carcinogenicity of chromium compounds. In: Nriagu JO, Nieboer E (ed) Chromium in the natural and human environments. John Wiley & Sons, New York, Chichester, Brisbane, Toronto, Singapore, pp 443–495
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Suzuki, Y., Fukuda, K. Reduction of hexavalent chromium by ascorbic acid and glutathione with special reference to the rat lung. Arch Toxicol 64, 169–176 (1990). https://doi.org/10.1007/BF02010721
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DOI: https://doi.org/10.1007/BF02010721