Cellular toxicity of hydrazine in primary rat hepatocytes.

Hydrazine (HzN) is an aircraft fuel and propellant used by the U.S. Air Force. The current study was undertaken to evaluate the acute toxicity of HzN in primary rat hepatocytes in vitro with reference to oxidative stress. The effects of short-term exposure (4 h) of hepatocytes to HzN were investigated with reference to viability, mitochondrial function, and biomarkers of oxidative stress. The viability data showed an increase in lactate dehydrogenase leakage and a decrease in mitochondrial activity with increasing concentration of HzN. The results of studies of oxidative stress biomarkers showed a depletion of reduced glutathione (GSH) and an increase in oxidized GSH, increased reactive oxygen species generation, lipid peroxidation, and reduced catalase activity. Furthermore, depletion of GSH and catalase activity in hepatocytes by buthionine sulfoximine and 3-amino triazole, respectively, prior to exposure to HzN, increased its toxicity. The results suggest that acute HzN-induced cytotoxicity in rat hepatocytes is likely to be mediated through oxidative stress.

[1]  L. Flohé,et al.  Assays of glutathione peroxidase. , 1984, Methods in enzymology.

[2]  Sten Orrenius,et al.  [4] Isolation and use of liver cells , 1978 .

[3]  T. Sendo,et al.  Metabolism and cytotoxicity of hydrazine in isolated rat hepatocytes. , 1987, Chemical & pharmaceutical bulletin.

[4]  C. Gosselin,et al.  Effect of hydrazine on urea cycle enzymes in vitro and in vivo. , 1971, Biochemical pharmacology.

[5]  M. Matsushita,et al.  Determination of proteins by a reverse biuret method combined with the copper-bathocuproine chelate reaction. , 1993, Clinica chimica acta; international journal of clinical chemistry.

[6]  J. Joseph,et al.  Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. , 1999, Free radical biology & medicine.

[7]  J. Timbrell,et al.  Identification and quantitation of hydrazine in the urine of patients treated with hydralazine , 1979, Clinical pharmacology and therapeutics.

[8]  R Doll,et al.  Occupational exposure to hydrazine and subsequent risk of cancer. , 1984, Occupational and environmental medicine.

[9]  B. Sinha.  Activation of hydrazine derivatives to free radicals in the perfused rat liver: a spin-trapping study. , 1987, Biochimica et biophysica acta.

[10]  H E Poulsen,et al.  Markers of oxidative damage to DNA: antioxidants and molecular damage. , 1999, Methods in enzymology.

[11]  J. Depierre,et al.  Hepatic oxidative stress and related defenses during treatment of mice with acetylsalicylic acid and other peroxisome proliferators. , 1995, Journal of biochemical toxicology.

[12]  R. Novak,et al.  Characterization of hydrazine-stimulated proteolysis in human erythrocytes. , 1988, Toxicology and applied pharmacology.

[13]  C. Cross,et al.  Free radicals, antioxidants, and human disease: where are we now? , 1992, The Journal of laboratory and clinical medicine.

[14]  D. D. Di Monte,et al.  Relationships between the mitochondrial transmembrane potential, ATP concentration, and cytotoxicity in isolated rat hepatocytes. , 1990, Archives of biochemistry and biophysics.

[15]  J. Timbrell,et al.  Studies on the Disposition and Metabolism of Hydrazine in Rats in vivo , 1992, Human & experimental toxicology.

[16]  H. Aebi,et al.  Catalase in vitro. , 1984, Methods in enzymology.

[17]  H. Kappus.  A survey of chemicals inducing lipid peroxidation in biological systems. , 1987, Chemistry and physics of lipids.

[18]  J. Timbrell,et al.  Effect of fructose on the biochemical toxicity of hydrazine in isolated rat hepatocytes. , 1997, Toxicology.

[19]  S. Kawanishi,et al.  Manganese-mediated oxidative damage of cellular and isolated DNA by isoniazid and related hydrazines: non-Fenton-type hydroxyl radical formation. , 1992, Biochemistry.

[20]  P. Seglen.  Preparation of isolated rat liver cells. , 1976, Methods in cell biology.

[21]  James B. Mitchell,et al.  Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. , 1987, Cancer research.

[22]  C. Dollery,et al.  Plasma Hydrazine Concentrations in Man after Isoniazid and Hydralazine Administration , 1985, Human toxicology.

[23]  J. Farber,et al.  Mechanisms of cell injury by activated oxygen species. , 1994, Environmental health perspectives.

[24]  G. Choudhary,et al.  Human health perspective on environmental exposure to hydrazines: a review. , 1998, Chemosphere.

[25]  B. P. Yu,et al.  Cellular defenses against damage from reactive oxygen species. , 1994, Physiological reviews.

[26]  E. Bredahl,et al.  Examination of the liver in personnel working with liquid rocket propellant , 1970, British journal of industrial medicine.

[27]  H. Sies,et al.  Glutathione and its role in cellular functions. , 1999, Free radical biology & medicine.

[28]  B. Mannervik,et al.  [59] Glutathione reductase , 1985 .

[29]  K. Yagi,et al.  Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. , 1979, Analytical biochemistry.

[30]  L. Colvin.  Metabolic fate of hydrazines and hydrazides. , 1969, Journal of pharmaceutical sciences.

[31]  J. Hayes,et al.  Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress. , 1999, Free radical research.

[32]  L. Witting.  Lipid peroxidationin vivo , 1965 .

[33]  Xiaodong Wang,et al.  Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c , 1996, Cell.

[34]  A. Nicolaou,et al.  Effect of hydrazine upon vitamin B12-dependent methionine synthase activity and the sulphur amino acid pathway in isolated rat hepatocytes. , 1999, Biochemical pharmacology.

[35]  Hydrazine radical formation catalyzed by rat microsomal NADPH-cytochrome P-450 reductase. , 1988, Biochemical and biophysical research communications.

[36]  J. Timbrell,et al.  Investigation of lipid peroxidation induced by hydrazine compounds in vivo in the rat. , 1989, Pharmacology & toxicology.

[37]  H. Söling,et al.  Inhibition of hepatic gluconeogenesis by phenethylhydrazine (phenelzine). , 1979, Biochemical pharmacology.

[38]  E. D. Harris,et al.  Regulation of antioxidant enzymes 1 , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[39]  S. Orrenius,et al.  Detoxification reactions in isolated hepatocytes. Role of glutathione peroxidase, catalase, and formaldehyde dehydrogenase in reactions relating to N-demethylation by the cytochrome P-450 system. , 1978, The Journal of biological chemistry.

[40]  W. Slikker,et al.  Reduced levels of catalase activity potentiate MPP+-induced toxicity: comparison between MN9D cells and CHO cells. , 1999, Toxicology letters.

[41]  R. Prough,et al.  Studies on the pathway of methane formation from procarbazine, a 2-methylbenzylhydrazine derivative, by rat liver microsomes. , 1983, Archives of biochemistry and biophysics.

[42]  J. Högberg,et al.  Glutathione depletion in isolated hepatocytes: its relation to lipid peroxidation and cell damage. , 2009, Acta pharmacologica et toxicologica.

[43]  D. Green,et al.  The Release of Cytochrome c from Mitochondria: A Primary Site for Bcl-2 Regulation of Apoptosis , 1997, Science.

[44]  R. C. Shank,et al.  Methylation of DNA guanine during the course of induction of liver cancer in hamsters by hydrazine or dimethylnitrosamine. , 1987, Carcinogenesis.

[45]  J. E. Willis.  The substitution of 1-methylhydrazine for ammonia in the glutamine synthetase system. , 1966, Biochemistry.

[46]  F. Tietze.  Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. , 1969, Analytical biochemistry.

[47]  M. Karbowski,et al.  Functional aspects of megamitochondria isolated from hydrazine‐ and ethanol‐treated rat livers , 2000, Pathology international.

[48]  Y. J. Kang,et al.  Suppression of Doxorubicin Cardiotoxicity by Overexpression of Catalase in the Heart of Transgenic Mice (*) , 1996, The Journal of Biological Chemistry.

[49]  N. DelRaso,et al.  EFFECT OF CULTURE CONDITIONS PRIOR TO EXPOSURE ON CADMIUM CYTOTOXICITY IN PRIMARY RAT HEPATOCYTES , 1999 .