ABSTRACT
In obesity, high levels of saturated fatty acids (SFAs) contribute to adipose tissue inflammation and dysfunction. Obesity-induced macrophage infiltration leads to insulin resistance, but the adipocyte itself may play a role in generating the inflammatory milieu. Given our recent findings of the role of TLR4 in myeloid biasing in obesity, we next investigated the role of TLR4 in adipocyte generated inflammatory responses to SFAs and lipopolysaccharides. We used WT and Tlr4-/- ear mesenchymal stem cell derived adipocytes (EMSC Ad) and bone marrow dendritic cells (BMDCs) to evaluate cell specific responses. Our work demonstrates a role for TLR4 in adipocyte- immune cell crosstalk and that SFA derived metabolites from adipocytes may induce proinflammatory stimulation of immune cells in a TLR4 independent manner.
Subject(s)
Adipocytes/metabolism , Disease Susceptibility , Inflammation/etiology , Inflammation/metabolism , Macrophages/metabolism , Toll-Like Receptor 4/metabolism , Adipogenesis , Adipose Tissue/metabolism , Adipose Tissue/pathology , Animals , Biomarkers , Cell Differentiation , Dendritic Cells/metabolism , Disease Models, Animal , Fatty Acids/metabolism , Gene Expression , Inflammation/pathology , Lipid Metabolism , Lipopolysaccharides/immunology , Macrophages/immunology , Male , Mice , Mice, Knockout , Toll-Like Receptor 4/geneticsABSTRACT
An in vitro activation assay for identifying suspected indirect mutagens and/or carcinogens is described. It involves the incubation of diffusion chambers filled with S-9 mix (S-9 fraction of rat liver and enzyme cofactors) and a compound under test with mammalian cells. The induction of sister chromatid exchanges (SCE) and/or chromosome aberrations can then be used as an indicator of mutagenicity or carcinogenicity. When a human lymphoid cell line, B35M, or Chinese hamster cell lines, V79 and CHO, are cultured with the diffusion chamber containing S-9 mix and the well-known indirect mutagen cyclophosphamide or its analog ifosfamide, a dose related increase in SCE or chromosome aberrations is observed in all the lines. The major advantages of this system over other in vitro activation assays are alleviation of cytotoxicity of S-9 mix and flexibility of treatment time.
Subject(s)
Biotransformation/drug effects , Chromosome Aberrations/chemically induced , Cyclophosphamide/pharmacology , Ifosfamide/pharmacology , Microsomes, Liver/drug effects , Sister Chromatid Exchange/drug effects , Animals , Antineoplastic Agents, Alkylating/pharmacology , Cells, Cultured , Cricetinae , Cricetulus , Crossing Over, Genetic , Diffusion Chambers, Culture , Humans , In Vitro Techniques , Male , Rats , Rats, Sprague-DawleyABSTRACT
The human genome holds an extraordinary trove of information about human development, physiology, medicine and evolution. Here we report the results of an international collaboration to produce and make freely available a draft sequence of the human genome. We also present an initial analysis of the data, describing some of the insights that can be gleaned from the sequence.