The potential of metabolomic approaches for investigating mode(s) of action of xenobiotics: case study with carbon tetrachloride.
ABSTRACT Both experimental animals and humans exhibit complex cellular responses upon exposure to xenobiotics and may undergo similar types of metabolic changes leading to adverse outcomes. Exposure to xenobiotics results in perturbation of many cellular events (e.g. oxidative stress, lipid peroxidation, inflammation, genotoxicity, cytotoxicity, etc.), and during this process biochemicals (endogenous metabolites) of a given metabolic pathway are increased, decreased or unaffected. Metabolomics is an emerging medium to high-throughput technology that can automatically identify, quantify and characterize hundreds to thousands of low molecular weight biochemicals simultaneously, using targeted or global analytical approaches, yielding a metabolic fingerprint and understanding of biochemical pathway perturbations. Herein, we illustrate how metabolomics can be utilized to explore the mechanisms of action of xenobiotics which affect different 'key events' contributing to different mode(s) of action. The extensively studied hepatotoxicant carbon tetrachloride (CCl(4)) is specifically described.
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ABSTRACT: Lycopene, the major carotenoid in tomatoes, is a known antioxidant that may lower oxidative stress biomarkers by a mechanism that is not fully elucidated. The intoxication of rats with carbon tetrachloride (CCl4) resulted in significant histological hepatic degradation accompanied by a marked increase in reactive oxygen species (ROS) and in the number of apoptotic cells. The induced oxidative stress in turn results in a significant elevation of lipid peroxidation and H2O2 generation, together with a decrease in the concentration of reduced glutathione (GSH) and a significant reduction in activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S transferase (GST). CCl4-intoxicated rats, pre-treated with lycopene, showed strongly reduced cell damage and ROS generation. The level of markers for hepatic integrity in lycopene pre-treated rats was close to the controls in the absence of CCl4 treatment, indicating the protective effect of lycopene pre-treatment. In the same way, lycopene pre-treated rats significantly increased SOD, CAT, GPx, GST activities and GSH level. In addition, we measured an increased lipoxygenase (LOX) activity in CCl4-intoxicated rats. This activity was reduced in lycopene pre-treated rats to values close to those observed in the controls, suggesting a potential pharmacological application of this dietary carotenoid.Journal of Functional Foods 10/2013; 5(4):1601–1610. · 4.48 Impact Factor
- Metabolomics 12/2014; 10(6):1293-1304. · 3.97 Impact Factor
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ABSTRACT: In this paper, we present an optimized procedure for metabolomic analysis of endogenous metabolites in mouse fibroblast (L929) cell line using gas chromatography/time-of-flight mass spectrometry with multivariate statistics. The optimization of metabolite extraction was performed using three solvents: methanol, water, and chloroform, and then followed by methoxymation and silylation. This method was subsequently validated using 29 reference standards and cell line samples. The intra- and inter-day relative standard deviations (RSDs) of the standard compounds were lower than 15.0 and 25.0 %, respectively. As for most of the tested metabolites in cell line samples, RSDs were below 20.0 % for reproducibility and stability, respectively. We applied this approach in metabolomic study of L929 cells obtained from TiO2 nanoparticle-induced cytotoxicity model samples (n = 5) and control samples (n = 5). Metabolite markers associated with TiO2 nanoparticle-induced cytotoxicity were identified and validated by statistical methods and reference standards. Our work highlights the potential of this method for cell metabolomic study.Chromatographia 11/2012; 75(21-22). · 1.37 Impact Factor