Analysis of the biological response of mouse liver (Mus musculus) exposed to As2O3 based on integrated -omics approaches

Abstract

Organic and inorganic mass spectrometries were used to investigate the biochemical response of mice (Mus musculus) to inorganic arsenic exposure using liver as the target organ. The toxicological effects of trivalent inorganic arsenic after oral administration (3 mg kg(-1) body weight and per day) were investigated over a period of 7 days using metallomics, metabonomics and redox proteomics approaches. Size-exclusion chromatography (SEC) with ICP-MS detection was combined with anion exchange chromatography (AEC) to characterize the biological response of the exposed mice. On the other hand, direct infusion mass spectrometry (DI-ESI-QTOF-MS) of polar and lipophilic extracts using positive and negative modes of acquisition (ESI+/ESI-) provided information about time-dependent changes in endogenous metabolites identified by Partial Least Square-Discriminant Analysis (PLS-DA). Finally, the study has been complemented with the evaluation of up/down-regulation of enzymes related to oxidative stress such as superoxide dismutase (SOD), glutathione reductase (GR), catalase (CAT) and peroxidases in connection with metal toxicity issues. The results show that the inorganic arsenic methylation in the liver may reach the saturation point upon chronic exposure to the element. On the other hand, SEC-ICP-MS coupling provided information about metal containing-proteins and metabolites related to arsenic exposure (metallomics) which has been correlated with the changes in the global metabolism (metabonomics), also considering their consequences on the redox status of protein and protein expression (redox proteomics). Our study shows that arsenic causes biochemical pathway alterations, such as energy metabolism (e.g. glycolysis, Krebs cycle), amino acid metabolism, choline metabolism and degradation of membrane phospholipids (apoptosis). This work illustrates the high reliability of the integrated use of organic mass spectrometry for the metabonomic study of biochemical effects induced by As2O3, with inorganic mass spectrometry for metallomic and speciation assessment of arsenic biomethylation in the liver of exposed mice, and redox proteomics to evaluate inhibition of enzymatic activity in different proteins such as superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) caused by this element. In conclusion, the integration of metallomics, metabolomics and redox proteomics results provides a more comprehensive evaluation about the biological response in experiments dealing with exposure to toxic metals.

Functional genomics and metabolomics reveal the toxicological effects of cadmium in Mus musculus mice

“…To acquire MS/MS spectra, N 2 was used as collision gas. Gas chromatography-mass spectrometry (GC-MS) analysis was also applied to mice plasma, as previously described (Garcia-Sevillano et al, 2013). Derivatizing agents, methoxylamine hydrochloride and N-methyl-N-(trimethylsilyl) trifluoroacetamide containing 1% trimethylchlorosilane, were obtained from Sigma-Aldrich.…”

Ochratoxin A induced early hepatotoxicity: new mechanistic insights from microRNA, mRNA and proteomic profiling studies

“…Through a cross-omics method, it is demonstrated that the major metabolic consequences of acute toxicity of aflatoxin B1, one of the well-known hepatotoxic mycotoxin, are associated with gluconeogenesis and lipid metabolism pathways 9 . Moreover, inorganic arsenic methylation in mouse liver may be saturated under chronic exposure to the element based on an integrated-omics approach 10 . The toxicity mechanism of another hepatotoxic compound EMD 335823 was also elucidated by using modern omics technologies, from which the liver pathologies are linked to deregulated PPARa signaling, glucose metabolism, and fatty acid metabolism 11 .…”

Combination of direct infusion mass spectrometry and gas chromatography mass spectrometry for toxicometabolomic study of red blood cells and serum of mice Mus musculus after mercury exposure

“…Despite the advantages of coupling separations techniques to mass spectrometry (e.g., matrix effects and isobaric interferences reduction), a number of studies recommend to use direct infusion of samples to mass spectrometer without any previous separation. This approach has been recently proposed in toxicological studies of mice [12,13,15,16] using high mass resolution analyzers, such as hybrid systems triple quadrupoletime-of-flight (QqQ-TOF). Under suitable experimental conditions, MS peak intensities are proportional to metabolite concentration and thus can be used to look for biomarkers related to toxic metal exposure.…”

“…Toxicometabolomics combines the use of: (i) experiment exposure to xenobiotics; (ii) multivariate pattern recognition based on metabolic profiling capabilities of multiple methodologies, including gas chromatography-mass spectrometry (GC-MS) [10], liquid chromatography-mass spectrometry (LC-MS) [11], direct infusion-mass spectrometry (DIMS) [11][12][13][14][15][16] and nuclear magnetic resonance (NMR) spectroscopy [8,9,17]; and (iii) chemometric analysis. Despite the advantages of coupling separations techniques to mass spectrometry (e.g., matrix effects and isobaric interferences reduction), a number of studies recommend to use direct infusion of samples to mass spectrometer without any previous separation.…”

“…The present work intends to determine the biochemical alterations of mouse metabolic pathways after 10-days subcutaneous injection of inorganic Hg, using mass spectrometry analysis followed by Partial Least Squares-Discrimination Analysis (PLS-DA) as a powerful tool to study the toxicological effects of xenobiotics in animal tissues and biological fluids [12,13,15,16]. Thus, DI-ESI(±)-QTOF-MS was applied to serum and RBCs of Hg-exposed Mus musculus mice to gather information about the effects of this element on metabolism of such cells.…”