PMID- 30071127 OWN - NLM STAT- MEDLINE DCOM- 20191004 LR - 20211204 IS - 1097-4652 (Electronic) IS - 0021-9541 (Linking) VI - 234 IP - 1 DP - 2018 Jan TI - Inflammation stimulates hypoxia-inducible factor-1alpha regulatory activity in 3T3-L1 adipocytes with conditioned medium from lipopolysaccharide-activated RAW 264.7 macrophages. PG - 550-560 LID - 10.1002/jcp.26763 [doi] AB - Obesity is a multifactorial, chronic, inflammatory disease that involves different processes, such as adipose tissue hypoxia. The aim of the current study was to characterize the effects of conditioned medium (CM) from lipopolysaccharide (LPS)-activated macrophages on the regulation of hypoxia-inducible factor 1alpha (HIF-1alpha)-related genes in murine adipocytes. For the in vitro analyses, 3T3-L1 murine adipocytes (9 days postdifferentiation) were incubated either in CM (25% medium of RAW 264.7 murine macrophages with 24 hr 500 ng/ml LPS), LPS at 500 ng/ml, or hypoxia (Hx; 1% O(2) , 94% N(2) , 5% CO(2) ) for 24 hr. For the in vivo experiments, mice were fed a high-fat diet. Both epididymal white adipose tissue (eWAT) and adipocytes in CM showed upregulation of Glut1, Mcp1, Il10, Tnf, and Il1b. The secretion of IL-6, TNF-alpha, and MCP-1 was also increased in CM-treated adipocytes. Moreover, increased levels of HIF-1alpha subunit and nuclear factor kappa B p65 were found after CM treatment, linking Hx, and inflammation. HIF-1alpha directly bound vascular endothelial growth factor A (Vegfa) and uncoupling protein 2 (Ucp2) genes, up- and downregulating its expression, respectively. Furthermore, the oxygen consumption rate was 30% lower in CM. The siRNA knockdown of mammalian target of rapamycin (Mtor) reversed the induction of HIF-1alpha found in CM. The macrophage infiltration simulated through CM seems to be a similar environment to an abnormally enlarged eWAT. We have evidenced that HIF-1alpha plays a regulatory role in the expression of Vegfa and Ucp2 in CM. Finally, the inhibition of the mTOR pathway prevented the HIF-1alpha activation induced by CM. The involvement of HIF-1alpha under proinflammatory conditions provides insight into the origins of Hx in obesity. CI - (c) 2018 Wiley Periodicals, Inc. FAU - Lopez-Pascual, Amaya AU - Lopez-Pascual A AUID- ORCID: 0000-0001-6091-8225 AD - Department of Nutrition, Food Science and Physiology, School of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain. AD - Centre for Nutrition Research, University of Navarra, School of Pharmacy and Nutrition, Pamplona, Spain. FAU - Lorente-Cebrian, Silvia AU - Lorente-Cebrian S AUID- ORCID: 0000-0002-1033-6152 AD - Department of Nutrition, Food Science and Physiology, School of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain. AD - Centre for Nutrition Research, University of Navarra, School of Pharmacy and Nutrition, Pamplona, Spain. AD - Nutrition Group, IdiSNA Navarra's Health Research Institute, Pamplona, Spain. FAU - Moreno-Aliaga, Maria J AU - Moreno-Aliaga MJ AUID- ORCID: 0000-0002-2018-6434 AD - Department of Nutrition, Food Science and Physiology, School of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain. AD - Centre for Nutrition Research, University of Navarra, School of Pharmacy and Nutrition, Pamplona, Spain. AD - Nutrition Group, IdiSNA Navarra's Health Research Institute, Pamplona, Spain. AD - Centre of Biomedical Research Network, CIBERobn Physiopathology of Obesity and Nutrition, ISCIII, Madrid, Spain. FAU - Martinez, J Alfredo AU - Martinez JA AUID- ORCID: 0000-0001-5218-6941 AD - Department of Nutrition, Food Science and Physiology, School of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain. AD - Centre for Nutrition Research, University of Navarra, School of Pharmacy and Nutrition, Pamplona, Spain. AD - Nutrition Group, IdiSNA Navarra's Health Research Institute, Pamplona, Spain. AD - Centre of Biomedical Research Network, CIBERobn Physiopathology of Obesity and Nutrition, ISCIII, Madrid, Spain. FAU - Gonzalez-Muniesa, Pedro AU - Gonzalez-Muniesa P AUID- ORCID: 0000-0002-3130-2496 AD - Department of Nutrition, Food Science and Physiology, School of Pharmacy and Nutrition, University of Navarra, Pamplona, Spain. AD - Centre for Nutrition Research, University of Navarra, School of Pharmacy and Nutrition, Pamplona, Spain. AD - Nutrition Group, IdiSNA Navarra's Health Research Institute, Pamplona, Spain. AD - Centre of Biomedical Research Network, CIBERobn Physiopathology of Obesity and Nutrition, ISCIII, Madrid, Spain. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180802 PL - United States TA - J Cell Physiol JT - Journal of cellular physiology JID - 0050222 RN - 0 (Ccl2 protein, mouse) RN - 0 (Chemokine CCL2) RN - 0 (Culture Media, Conditioned) RN - 0 (Glucose Transporter Type 1) RN - 0 (Hif1a protein, mouse) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (Interleukin-6) RN - 0 (Lipopolysaccharides) RN - 0 (Slc2a1 protein, mouse) RN - 0 (Tumor Necrosis Factor-alpha) RN - 0 (Uncoupling Protein 2) RN - 0 (Vascular Endothelial Growth Factor A) RN - 0 (vascular endothelial growth factor A, mouse) RN - EC 2.7.1.1 (mTOR protein, mouse) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - 3T3-L1 Cells MH - Adipocytes/drug effects MH - Adipose Tissue, White/drug effects/metabolism MH - Animals MH - Chemokine CCL2/genetics MH - Culture Media, Conditioned/pharmacology MH - Glucose Transporter Type 1/genetics MH - Humans MH - Hypoxia-Inducible Factor 1, alpha Subunit/*genetics MH - Inflammation/*genetics/pathology MH - Interleukin-6/genetics MH - Lipopolysaccharides/pharmacology MH - Macrophages/drug effects/metabolism MH - Mice MH - RAW 264.7 Cells MH - TOR Serine-Threonine Kinases/genetics MH - Tumor Necrosis Factor-alpha/genetics MH - Uncoupling Protein 2/*genetics MH - Vascular Endothelial Growth Factor A/*genetics OTO - NOTNLM OT - anaerobic OT - coculture OT - diet-induced obesity (DIO) OT - in vitro model OT - obese phenotype OT - oxygenation EDAT- 2018/08/03 06:00 MHDA- 2019/10/08 06:00 CRDT- 2018/08/03 06:00 PHST- 2018/01/30 00:00 [received] PHST- 2018/04/27 00:00 [accepted] PHST- 2018/08/03 06:00 [pubmed] PHST- 2019/10/08 06:00 [medline] PHST- 2018/08/03 06:00 [entrez] AID - 10.1002/jcp.26763 [doi] PST - ppublish SO - J Cell Physiol. 2018 Jan;234(1):550-560. doi: 10.1002/jcp.26763. Epub 2018 Aug 2.