RESUMO
Hypomethylation of DNA repeats has been linked to diseases and cancer predisposition. Human studies suggest that higher blood concentrations of environmental contaminants (EC) correlate with levels of hypomethylation of DNA repeats in blood. The objective of this study was to examine the effect of in utero and/or lactational exposure to EC on the methylation of DNA repeats (LINE-1 and identifier element) in Sprague-Dawley rat pups at birth, at postnatal day (PND) 21, and in adulthood (PND78-86). From gestation day 0 to PND20, dams were exposed to a mixture "M" of polychlorinated biphenyls (PCB), pesticides, and methylmercury (MeHg), at 0.5 or 1 mg/kg/d (0.5M and M). At birth, some control (C) and M litters were cross-fostered to create the following in utero/postnatal exposure groups: C/C, M/C, C/M, M/M. Additional dams received 1.8 ng/kg/d of a mixture of aryl-hydrocarbon receptor (AhR) agonists (non-ortho-PCB, PC-dibenzodioxins, and PC-dibenzofurans) without or with 0.5M (0.5MAhR). Measurements of EC residue levels confirmed differences in their accumulation across treatments, age, and tissues. Although induction of hepatic detoxification enzyme activities (cytochrome P-450) demonstrated biological effects of treatments, the assessment of methylation in DNA repeats by sodium bisulfite pyrosequencing of liver, spleen, and thymus samples revealed no marked treatment-related effects but significant tissue- and age-related methylation differences. Further studies are required to determine whether absence of significant observable treatment effects on methylation of DNA repeats in the rat relate to tissue, strain, or species differences.
Assuntos
Metilação de DNA/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Lactação , Elementos Nucleotídeos Longos e Dispersos/efeitos dos fármacos , Exposição Materna/efeitos adversos , Animais , Feminino , Masculino , Compostos de Metilmercúrio/toxicidade , Praguicidas/toxicidade , Bifenilos Policlorados/toxicidade , Gravidez , Ratos , Ratos Sprague-Dawley , Análise de Sequência de DNA , Sulfitos/químicaRESUMO
Non-alcoholic fatty liver disease (NAFLD), defined by the American Liver Society as the buildup of extra fat in liver cells that is not caused by alcohol, is the most common liver disease in North America. Obesity and type 2 diabetes are viewed as the major causes of NAFLD. Environmental contaminants have also been implicated in the development of NAFLD. Northern populations are exposed to a myriad of persistent organic pollutants including polychlorinated biphenyls, organochlorine pesticides, flame retardants, and toxic metals, while also affected by higher rates of obesity and alcohol abuse compared to the rest of Canada. In this study, we examined the impact of a mixture of 22 contaminants detected in Inuit blood on the development and progression of NAFLD in obese JCR rats with or without co-exposure to 10% ethanol. Hepatosteatosis was found in obese rat liver, which was worsened by exposure to 10% ethanol. NCM treatment increased the number of macrovesicular lipid droplets, total lipid contents, portion of mono- and polyunsaturated fatty acids in the liver. This was complemented by an increase in hepatic total cholesterol and cholesterol ester levels which was associated with changes in the expression of genes and proteins involved in lipid metabolism and transport. In addition, NCM treatment increased cytochrome P450 2E1 protein expression and decreased ubiquinone pool, and mitochondrial ATP synthase subunit ATP5A and Complex IV activity. Despite the changes in mitochondrial physiology, hepatic ATP levels were maintained high in NCM-treated versus control rats. This was due to a decrease in ATP utilization and an increase in creatine kinase activity. Collectively, our results suggest that NCM treatment decreases hepatic cholesterol export, possibly also increases cholesterol uptake from circulation, and promotes lipid accumulation and alters ATP homeostasis which exacerbates the existing hepatic steatosis in genetically obese JCR rats with or without co-exposure to ethanol.
Assuntos
Poluentes Ambientais/efeitos adversos , Fígado Gorduroso/induzido quimicamente , Metabolismo dos Lipídeos/efeitos dos fármacos , Trifosfato de Adenosina/metabolismo , Animais , Colesterol/metabolismo , Exposição Ambiental , Etanol/toxicidade , Homeostase/efeitos dos fármacos , Humanos , Inuíte , Fígado/efeitos dos fármacos , Fígado/metabolismo , Ratos EndogâmicosRESUMO
BACKGROUND: The geographic distribution of environmental toxins is generally not uniform, with certain northern regions showing a particularly high concentration of pesticides, heavy metals and persistent organic pollutants. For instance, Northern Canadians are exposed to high levels of persistent organic pollutants like polychlorinated biphenyls (PCB), organochlorine pesticides (OCs) and methylmercury (MeHg), primarily through country foods. Previous studies have reported associations between neuronal pathology and exposure to such toxins. The present investigation assessed whether perinatal exposure (gestation and lactation) of rats to a chemical mixture (27 constituents comprised of PCBs, OCs and MeHg) based on Arctic maternal exposure profiles at concentrations near human exposure levels, would affect brain levels of several inflammatory cytokines METHODS: Rats were dosed during gestation and lactation and cytokine levels were measured in the brains of offspring at five months of age. Hypothalamic cytokine protein levels were measured with a suspension-based array system and differences were determined using ANOVA and post hoc statistical tests. RESULTS: The early life PCB treatment alone significantly elevated hypothalamic interleukin-6 (IL-6) levels in rats at five months of age to a degree comparable to that of the entire chemical mixture. Similarly, the full mixture (and to a lesser degree PCBs alone) elevated levels of the pro-inflammatory cytokine, IL-1b, as well as the anti-inflammatory cytokine, IL-10. The full mixture of chemicals also moderately increased (in an additive fashion) hypothalamic levels of the pro-inflammatory cytokines, IL-12 and tumor necrosis factor (TNF-α). Challenge with bacterial endotoxin at adulthood generally increased hypothalamic levels to such a degree that differences between the perinatally treated chemical groups were no longer detectable. CONCLUSIONS: These data suggest that exposure at critical neurodevelopmental times to environmental chemicals at concentrations and combinations reflective of those observed in vulnerable population can have enduring consequences upon cytokines that are thought to contribute to a range of pathological states. In particular, such protracted alterations in the cytokine balance within the hypothalamus would be expected to favor marked changes in neuro-immune and hormonal communication that could have profound behavioral consequences.
Assuntos
Poluentes Ambientais/toxicidade , Hidrocarbonetos Clorados/toxicidade , Hipotálamo/efeitos dos fármacos , Hipotálamo/crescimento & desenvolvimento , Compostos de Metilmercúrio/toxicidade , Bifenilos Policlorados/toxicidade , Efeitos Tardios da Exposição Pré-Natal/imunologia , Análise de Variância , Animais , Regiões Árticas , Carga Corporal (Radioterapia) , Canadá , Modelos Animais de Doenças , Poluentes Ambientais/farmacologia , Feminino , Hipotálamo/imunologia , Hipotálamo/metabolismo , Interleucinas/análise , Interleucinas/imunologia , Lactação , Lipopolissacarídeos/imunologia , Exposição Materna , Gravidez , Complicações na Gravidez , Ratos , Tempo , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Analysis of alkoxyacetic acids has received considerable research interest in toxicology because these compounds have been reported as metabolites and biomarkers of exposure to widely used industrial chemicals such as alkyl-substituted ethylene glycols and other aliphatic ethers. This paper describes an improved method for the determination of methoxyacetic acid (MAA), ethoxyacetic acid (EAA), and butoxyacetic acid (BAA) in rat urine. Solid-phase extraction with Bakerbond(T) C18 bonded silica cartridges was successfully employed to isolate the acids from rat urine. The acids were then converted to methyl esters with diazomethane derivatization and analyzed using a gas chromatograph (GC) equipped with a mass spectrometry (MS) and a GC with flame ionization detector (FID). Employing GC-MS under selected ion monitoring detection, the lowest detection concentrations for MAA, EAA, and BAA were determined to be from 2 to 4 ng/mL urine in 1 mL of sample size. This method is 5 to 10 times more sensitive than that using GC-FID. The method described here is superior to the existing ones reported in the literature in that it employs an easy sample treatment procedure and gives much higher recoveries, making it suitable for routine assays. The utility of this new method was demonstrated in a toxicology study of aliphatic alkyl ethers.