Antidotal Efficacy of Antioxidants against Cyanide Poisoning in vitro.
AbstractCyanide is a potent homicidal, genocidal and chemical warfare agent. Besides, its known inhibitory effects on various enzyme Systems, its other pronounced toxic effects include lipid peroxidation (LPx), particularly in the central nervous system or neuronal cells in vitro. The present study assessed the cytotoxicity of potassium cyanide (KCN) in two non-neuronal mammalian cell cultures, viz., human embryonic lung epithelium (L-132) and baby hamster kidney (BHK-21 ) cells. In addition, the cytoprotective potential of two antioxidant agents, namely, curcumin (CMN) and N-acetylcysteine (NAC) against KCN (2 and 4 mM) in vitro was evaluated. In both the cell lines, KCN reduced cell viability as indicated by trypan blue dye exclusion, leakage of cytosolic lactate dehydrogenase and neutral red uptake. Protein content was unaffected in L-132 cells while cellular respiration determined by MTT assay) was impaired in both the cells. A dose-dependent glutathione mediated LPx was observed in BHK-21 cells alone. The above cytotoxic changes produced by KCN were more effectively minimised by NAC as compared to CMN. Efficacy of CMN and NAC have therapeutic implications as adjuncts to existing cyanide antidotes.
Way, J.L. Cyanide intoxication and its mechanism of antagonism. Ann. Rev. Pharmacol., 1984,24,451-81.
Johnson, J.D.; Conroy, W.G.; Bums, K.D. & Isom, G.E. Peroxidation of brain lipids following cyanide intoxication in mice. Toxicology, 1987, 46,2 1-28.
Ardelt, B.K.; Borowitz, J.L. & Isom, G.E. Brain lipid peroxidation and antioxidant protectant mechanisms following acute cyanide intoxication. Toxicology, 1989, 56, 147-54.
Ardelt, B.K.; Borowitz, J.L.; Maduh, E.U.; Swain, S.L. & Isom, G.E. Cyanide-induced lipid peroxidation in different organs: Subcellular distribution and hydroperoxide generation in neuronal cells. Toxicology, 1994,89, 127
Sharma, S.C.; Mukhtar, H.; Sharma, S.C. & Krishna Murti, C.R. Lipid peroxide formation in experimental inflammation. Biochemistry Pharmacology, l972,21, 1210-14.
Soudamini, K.K.; Unnikrishnan, M.C.; Soni, K.B. & Kuttan, R. Inhibition of lipid peroxidation and cholesterol levels in mice by curcumin. Indian J. Physiol. Pharmacol., 1 992,36, 239-43.
Sharma, O.P. Antioxidant activity of curcumin and related compounds. Biochemistry Pharmacology, 1976,25, 1 8 1 1 - 12.
Henderson, A. & Hayes, P. Acetylcysteine as a cytoprotective antioxidant in patients with severe sepsis: Potential new use for an old drug. Critical Care, 1994,28, 1086-88.
Ballantyne, B. Toxicology of cyanides. In Clinical and experimental toxicology of cyanides, edited by B. Ballantyne and T.C. Marrs. Wright Publishers, Bristol, UK, 1987. pp 112.
Borenfreund, E.; Babich, H. & Martin-Alguacil, N. Comparison of two in vitro cytotoxicity assays the neutral red (NR) and tetrazolium MTT tests. Toxicology in Vitro, 1988,2, 1-6.
Mosmann, T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol.Methods, 1983, 65, 55-63.
Bradford, M.M. A rapid and sensitive method for quantification of microgram quantities of protein utilising the principle of protein dye binding. Analytical Biochemistry, 1 976, 72,248-54.
Ellman, G.L. Tissue sulfhydryl groups. Arch. Biochem. Biophys., 1959,82,70-77.
Buege, J.A. & Aust, S.D. Microsomal lipid peroxidation. Methods in Enzymology., 1 978,52, 302.
Gunasekar, P.G.; Borowitz, J.L. & Isom, G.E. Cyanide stimulates reactive oxygen species generation through the cyclo-oxygenase lipo-oxygenase pathways: Implications for cytotoxicity. The Toxicologist, 1996,30,98.
Husoy, T.; Syversen, T. & Jenssen, J. Comparison of four in vitro cytotoxicity tests: The MTT assay, NR assay, Uridine incorporation and protein measurement. Toxicology in vitro, 1993,7, 149-54.
Trump, B.F. & Berezesky, I.K. Cellular and molecular pathobiology of reversible and irreversible injury. In vitro toxicity indicators, edited by C.A. Tyson and J.M. Frazier (Methods in Enzymology). Academic Press Inc.,
California, USA, 1994. pp 7-9.
Ponsoda, X.; Jover, R.; Castell, J.V. & Gomez-Lechon, M.J. In vitro toxicity to two cellular systems of first ten chemicals on the MElC list. ATLA, 1990,17,2 18-23.
Ercal, N.; Teeratphan, P.; Lutz, P.; Hammond T.C. & Matthews, R.H. N-acetylcysteine protects chinese hamster ovary (CHO) cells 'from lead-induced oxidative stress. Toxicology, 1996, 108,57-64.
Piper, J.T.; Awasthi, S.; Chaubey, M.; Singhal. S.S.; Srivastava, S.K. & Awasthi, Y.C. Effect of curcumin on glutathione (GSH) levels and GSH-related enzymes in rat liver. The Toxicologist, 1996,30,285-86.
Pulla Reddy, A. Ch. & Lokesh, B.R. Effect of curcumin and eugenol on iron-induced hepatic toxicity in rats. Toxicology, 1996, 107,39-45.
Where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India