Liver toxicity induced by munitions compounds TNT, 2,6-DNT, 2,4-DNT, 4A-DNT, and 2A-DNT. Rattus norvegicus

The use of a systems biology approach to analyze common and specific mechanisms of liver toxicity induced by munitions compounds TNT, 2,6-DNT, 2,4-DNT, 4A-DNT, and 2A-DNT The munitions compound 2,4,6-trinitrotoluene (TNT), its environmental degradation products 2-amino-4,6-dinitrotoluene (2A-DNT) and 4-amino-2,6-dinitrotoulene (4A-DNT), and two other munitions, 2,4-dinitrotoluene (2,4-DNT) and 2,4-dinitrotoluene (2,6-DNT) contaminate contaminate land, water and retired ammunitions plants. The release of these compounds to the environment is due to military activities and a series of manufacturing processes. Although toxicity has been characterized for these compounds, little is known of their mechanism of action. Here we describe to use an integrative systems biology approach including toxicology, pathology, transcriptomics, metabolomics, gene function classification, pathway analysis and gene network modeling to try to understand the mechanisms of toxicity of these compounds. Overall design: Changes in gene expression were tested using Agilent commercial whole rat genome microarrays (4 X 44K). Experiments were designed for both purposes of temporal and dose response analysis. For temporal analysis, only two time points for each compound were used, 24h and 48h. For dose response analysis, four doses plus a vehicle control were employed for each compound at each time point. A separate control was used for different compounds even at the same time point. The dose selection was based on the LD 50 data for each compound. Except controls, the lowest dose, the second lowest dose, the second highest dose and the highest dose were equivalent to respectively for each compound (Table 1). Four biological replicates of this design were conducted, each using different animals, and four biological replicates were applied, which resulted in a total of 200 microarrays. ( one array was deleted from the set, 199 arrays used)