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Effects of 1,3,5-Trinitrobenzene on Cytotoxicity and Metabolic Activity of Type I Astrocytes of Rats

Department of Pathology, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma, USA

Gunda Reddy

Directorate of Toxicology, U.S. Army Center for Health Promotion and Preventive Medicine, Aberdeen Proving Ground, Maryland, USA

Jerry W. Ritchey

Department of Pathology, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma, USA

Jeremiah T. Saliki

Oklahoma Animal Disease Diagnostic Laboratory, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma, USA

Charles W. Qualls, Jr.

Department of Pathology, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma, USA

Correspondence: Address correspondence to Gunda Reddy, PhD, DABT, Toxicologist, U.S. Army Center for Health Promotion and Preventive Medicine, 5158 Blackhawk Road, Aberdeen Proving ground, MD 21010-5403, USA. E-mail:gunda.reddy{at}apg.amedd.army.mil

1,3,5-Trinitrobenzene (TNB) is a munitions chemical that causes gliovascular lesions in the brain stem of rats similar to those produced by thiamine deficiency and nitroaromatic compounds, including m-dinitrobenzene. To identify neuropathic indices of toxicity, the effects of varying concentrations (0 to 2 mM) of TNB on cytotoxicity and cellular metabolic activity were examined using cultured astrocytes from Fischer-344 rats. The cytotoxicity was assessed by lactate dehydrogenase (LDH) leakage into the culture medium. Astrocyte metabolic activity was assessed by measuring the conversion of a tetrazolium salt to a formazan product. Additionally, the effects of oxidative stress on cellular metabolic activity - were determined by varying oxygen tension via alteration of culture media depth. In vitro, the toxic concentration 50% (TC₅₀) of TNB, which induced cell death, was 16 µM following a 24-h exposure. The concentration of TNB that reduced cellular metabolic activity by 50% was 29 µM following a 24-h exposure. Varying the depth of the culture media did not influence the cellular metabolic activity in control or TNB-treated astrocytes. These results support the hypothesis that TNB induced neurotoxicity could partially be mediated via injury to astrocytes, a major component of the blood-brain barrier.

Key Words: Astrocytes • Cytotoxicity • Metabolic Activity • Rats, 1,3,5-Trinitrobenzene

International Journal of Toxicology, Vol. 24, No. 1, 51-57 (2005)
DOI: 10.1080/10915810590918724


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