Abstract
Previous studies have demonstrated that an aqueous smokeless tobacco extract (STE) administered in an acute oral dose to rats induces an enhanced induction of hepatic mitochondrial and microsomal lipid peroxidation, hepatic nuclear DNA single strand breaks, enhanced excretion of urinary lipid metabolites, including malondialdehyde, formaldehyde, acetaldehyde and acetone, and increased production of nitric oxide (NO) by peritoneal macrophage cells. These observations indicate that STE induces the production of oxygen free radicals. We have therefore examined the in vitro incubation of cultured J774A.1 macrophage cells with STE on the release of the enzyme lactate dehydrogenase (LDH) into the media as an indicator of cellular membrane damage and cytotoxicity. The amount of LDH released by STE was both concentration- and time-dependent. The cytotoxicity of STE to macrophage J774A.1 cells in culture was further determined from percent viability after various periods of incubation. The addition of 250 μg STE/ml to the cultured J774A.1 cells resulted in a 2.9-fold increase in the release of LDH. Individual coincubation with superoxide dismutase (SOD), catalase, mannitol, and allopurinol had no significant effect on the release of LDH into the culture medium, while a combination of the four free radical scavengers resulted in a 59% decrease in the STE-induced release of LDH. At 75 μM concentrations of viramine E and vitamin E succinate, approximately 28% and 41% inhibitions were observed in STE-induced LDH leakage, respectively. Taken together with previous studies, the results indicate that STE activates macrophage cells, resulting in the production of reactive oxygen species. These oxygen free radicals may be responsible for tissue damaging effects including membrane damage, and selected oxygen free radical scavengers and antioxidants can attenuate these tissue damaging effects.
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Bagchi D, Hassoun EA, Bagchi M, Stohs SJ (1993) Protective effects of antioxidants against endrin-induced hepatic lipid peroxidation, DNA damage, and excretion of urinary lipid metabolites. Free Rad Biol Med 15:217–222
Bagchi M, Bagchi D, Hassoun EA, Stohs SJ (1994) Smokeless tobacco induced increases in hepatic lipid peroxidation, DNA damage, and excretion of urinary lipid metabolites. Int J Exptl Pathol 75:197–202
Bagchi M, Hassoun E, Akubue P, Bagchi D, Stohs SJ (1993) Comparative effects of endrin on hepatic lipid peroxidation and DNA damage, and nitric oxide production by peritoneal macrophages from C57BL/6J and DBA/2 mice. Comp Biochem Physiol 105C:525–529
Brinton LA, Blot WJ, Becker JA, Winn DM, Browder JP, Framer JC Jr, Frauneni JF Jr (1994) A case control study of cancers of the nasal cavity and paranasal sinuses. Am J Epidemiol 119:896–906
Butler AR, Williams DLH (1993) The physiological role of nitric oxide. Chem Soc Rev 22:233–241
Christen AG, McDaniel RK, McDonald JR Jr (1990) The smokeless tobacco ‘time bomb’. Postgrad Med 87:69–74
Fielding JE (1985) Smoking: Health effects and controls (First of two parts). New Engl J Med 313:491–498
Hassoun EA, Bagchi D, Bagchi M, Stohs SJ (1995) Effect of vitamin E succinate on smokeless tobacco-induced production of nitric oxide by rat peritoneal macrophages and J774A.1 macrophage cells in culture. Free Rad Biol Med 18:577–583
Hassoun EA, Roche VF, Stohs SJ (1993) Release of enzymes by ricin from macrophages and Chinese hamster ovary cells in culture. Toxicol Meth 3:119–129
Hecht SS, Rivenson A, Barley J, DiBello J, Adams JD, Hoffmann D (1986) Induction of oral cavity tumors in F344 rats by tobaccospecific nitrosamines and snuff. Cancer Res 46:4162–4166
Johansson SL, Hirsch JH, Larsson PA, Sadi J, Osterdahl BG (1989) Snuff-induced carcinogenesis: Effect of snuff in rats initiated with 4-nitroquinoline-N-oxide. Cancer Res 49:3063–3069
Kalra J, Chaudhury AK, Prasad K (1991) Increased production of oxygen free radicals in cigarette smokers. Int J Exptl Pathol 72:1–7
Klee S, Nurnberger MC, Ungemach FR (1994) The consequences of nitrofurantoin-induced oxidative stress in isolated hepatocytes: Evaluation of pathobiochemical alterations. Chem-Biol Interact 93:91–102
Moss DW, Henderson AR, Kachmar JR (1986) Enzymes. In: Tietz (ed) Textbook of Clinical Chemistry. Saunders Co, Philadelphia, pp 619–763
Nathan C (1992) Nitric oxide as a secretory product of mammalian cells. FASEB J 6:3051–3064
Stich HF, Anders F (1989) The involvement of reactive oxygen species in oral cancers of betel quid/tobacco chewers. Mutat Res 214:47–61
Stohs SJ, Bagchi M, Bagchi D, Hassoun EA (1994) In vitro and in vivo induction of oxidative stress by smokeless tobacco extract (STE). Toxicologist 13:336
Stuehr DJ, Nathan CF (1989) A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med 169:1543–1555
US Surgeon General (1986) The health consequences of using smokeless tobacco. NIH publication no. 862-2874, Washington, DC
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Bagchi, D., Hassoun, E.A., Bagchi, M. et al. Protective effects of free radical scavengers and antioxidants against smokeless tobacco extract (STE)-induced oxidative stress in macrophage J774A.1 cell cultures. Arch. Environ. Contam. Toxicol. 29, 424–428 (1995). https://doi.org/10.1007/BF00212511
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DOI: https://doi.org/10.1007/BF00212511