After scratching a gap into confluent monolayers, we followed gap closure by live cell imaging

in disorders where the cross-talk between inflammation and lipid metabolism is essential to the development of pathogenesis, as it is in diet-induced metabolic syndrome. the activation of pparc may inhibit nf-kb and therefore suppress inflammation, but in turn evoke transition to pathological state, in this case hepatic steatosis. despite the apparent harmful effects of inflammation, such as triggering insulin resistance, its protective physiological role in preventing transitions to even less preferable system states should not be neglected. thus, the tradeoff between the beneficial and harmful effects of altered pparc activity should be carefully considered when using pparc ligands and nf-kbinhibiting agents to ameliorate metabolic syndrome associated pathologies. the presented findings demonstrate the use of highthroughput dataset analyses as a starting point for generating testable hypotheses that may open new avenues for dietary prevention strategies, clinical research and pharmaceutical therapies. Materials and Methods Ethics Statement Animal experiments were approved by the Institutional Animal Care and Use Committee of the Netherlands Organization for Applied Scientific Research and were in compliance with European purchase Tedizolid (phosphate) Community specifications regarding the use of laboratory animals. Animals and diets The study involved 186 male ApolipoproteinE3-Leiden transgenic mice at 1462 weeks of age. Apolipoprotein E3-Leiden Hepatic Effects of HF Diets transgenic mice display lipoprotein profile that closely resembles that of humans and develop human-like dysbetalipoproteinemia and atherosclerotic lesions when fed Western-type diets. The age of 14 weeks was chosen as optimal because the animals are considered adult at that stage. The first group of ApoE3L mice was fed standard chow diet DU; Special Diet Services, Witham, Essex, UK), the second group of mice was fed a high-fat diet based on animal fats and the third group of mice was fed a high-fat diet based on plant fats. The macronutrient content and the fatty acid composition of chow and high-fat diets are provided in Supporting Experimental design and sample preparation From three weeks prior to 17984313 diet intervention onwards, all animals were fed a standard chow diet. At the beginning of the study, mice were divided into three groups: control group fed chow diet, group fed HFBT diet and group fed HFP diet. Because the interest of the study was to asses effects of high-fat diets 11325787 under physiological conditions, animals were fed ad libitum. Series of control experiments employing a metabolic cage setup showed that C57Bl/6 mice, the genetic background of the APOE3L mice, have isocaloric food intake when fed low fat and HFBT and HFP diets. The light cycles were identical for all animals. For mRNA expression profiling, six mice from each diet group were sacrificed at time points 0 days, 1 day, 3 days, 1, 2, 4, 8, 12 and 16 weeks, their livers were dissected after 4 hour fasting period, snap frozen in liquid nitrogen and stored at 280uC until further processing. RNA isolation, labeling and hybridization to microarrays Total RNA was isolated using TRIzol reagent according to the manufacturer’s instructions. RNA was treated with DNAse and purified using the SV total RNA isolation system. Concentrations and purity of RNA samples were determined on a NanoDrop ND1000 spectrophotometer. RNA integrity was checked on an Agilent 2100 bioanalyzer with 6000 Nano Chips according to the manufacturer’s instruction

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