The Effect of Pericellular Oxygen Levels on Proteomic Profile and Lipogenesis in 3T3-L1 Differentiated Preadipocytes Cultured on Gas-Permeable Cultureware

. 2016 ; 11 (3) : e0152382. [epub] 20160329

Jazyk angličtina Země Spojené státy americké Médium electronic-ecollection

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/pmid27023342

Pericellular oxygen concentration represents an important factor in the regulation of cell functions, including cell differentiation, growth and mitochondrial energy metabolism. Hypoxia in adipose tissue has been associated with altered adipokine secretion profile and suggested as a possible factor in the development of type 2 diabetes. In vitro experiments provide an indispensable tool in metabolic research, however, physical laws of gas diffusion make prolonged exposure of adherent cells to desired pericellular O2 concentrations questionable. The aim of this study was to investigate the direct effect of various O2 levels (1%, 4% and 20% O2) on the proteomic profile and triglyceride accumulation in 3T3-L1 differentiated preadipocytes using gas-permeable cultureware. Following differentiation of cells under desired pericellular O2 concentrations, cell lysates were subjected to two-dimensional gel electrophoresis and protein visualization using Coomassie blue staining. Spots showing differential expression under hypoxia were analyzed using matrix-assisted laser desorption/ionization mass spectrometry. All identified proteins were subjected to pathway analysis. We observed that protein expression of 26 spots was reproducibly affected by 4% and 1% O2 (17 upregulated and 9 downregulated). Pathway analysis showed that mitochondrial energy metabolism and triglyceride synthesis were significantly upregulated by hypoxia. In conclusion, this study demonstrated the direct effects of pericellular O2 levels on adipocyte energy metabolism and triglyceride synthesis, probably mediated through the reversed tricarboxylic acid cycle flux.

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Rizzatti V, Boschi F, Pedrotti M, Zoico E, Sbarbati A, Zamboni M. Lipid droplets characterization in adipocyte differentiated 3T3-L1 cells: size and optical density distribution. Eur J Histochem. 2013;57: 24 10.4081/ejh.2013.e24 PubMed DOI PMC

Lastra G, Sowers JR. Obesity and cardiovascular disease: role of adipose tissue, inflammation, and the renin-angiotensin-aldosterone system. HormMolBiolClinInvestig. 2013;15: 49–57. PubMed

Song X, Jousilahti P, Stehouwer CD, Soderberg S, Onat A, Laatikainen T, et al. Cardiovascular and all-cause mortality in relation to various anthropometric measures of obesity in Europeans. NutrMetab Cardiovasc. Department of Public Health, Hjelt Institute, University of Helsinki, Helsinki, Finland; Department of Chronic Disease Prevention, National Institute for Health and Welfare, Helsinki, Finland. Electronic address: xin.song@helsinki.fi Department of Chronic; 2014;

Carmichael AR. Obesity as a risk factor for development and poor prognosis of breast cancer. BJOG. Department of Surgery, Russells Hall Hospital, Dudley, UK. homepac@doctors.org.uk; 2006;113: 1160–1166. PubMed

Hong S, Cai Q, Chen D, Zhu W, Huang W, Li Z. Abdominal obesity and the risk of colorectal adenoma: a meta-analysis of observational studies. EurJCancer Prev. Department of Gastroenterology, Changhai Hospital of Second Military Medical University, Shanghai, China; 2012;21: 523–531. PubMed

Horikoshi T, Balin AK, Carter DM. Effect of oxygen on the growth of human epidermal keratinocytes. J Invest Dermatol. 1986;86: 424–427. PubMed

Horikoshi T, Balin AK, Carter DM. Effects of oxygen tension on the growth and pigmentation of normal human melanocytes. J Invest Dermatol. 1991;96: 841–844. PubMed

Balin AK, Pratt L. Oxygen modulates the growth of skin fibroblasts. In Vitro Cell Dev Biol Anim. 2002;38: 305–310. 10.1290/1071-2690(2002)038<0305:OMTGOS>2.0.CO;2 PubMed DOI

Semenza GL. Regulation of Metabolism by Hypoxia-Inducible Factor 1. Cold Spring HarbSympQuantBiol. Vascular Program, Institute for Cell Engineering, Departments of Pediatrics, Medicine, Oncology, Radiation Oncology, and Biological Chemistry, and McKusick-Nathans Institute of Genetic Medicine, The Johns Hopkins University School of Medicine, Baltimore,; 2011;

Bambrick LL, Kostov Y, Rao G. In vitro cell culture pO2 is significantly different from incubator pO2. Biotechnol Prog. 2011;27: 1185–1189. 10.1002/btpr.622 PubMed DOI

Metzen E, Wolff M, Fandrey J, Jelkmann W. Pericellular PO2 and O2 consumption in monolayer cell cultures. RespirPhysiol. Physiologisches Institut I, Rheinische Friedrich-Wilhelms-Universitat, Bonn, Germany; 1995;100: 101–106. 10.1016/0034-5687(94)00125-J PubMed DOI

Allen CB, Schneider BK, White CW. Limitations to oxygen diffusion and equilibration in in vitro cell exposure systems in hyperoxia and hypoxia. AmJPhysiol Lung Cell MolPhysiol. Department of Pediatrics, National Jewish Medical and Research Center, Denver, Colorado 80206, USA. allenc@njc.org; 2001;281: L1021–L1027. PubMed

Trayhurn P. Hypoxia and adipose tissue function and dysfunction in obesity. Physiol Rev. 2013;93: 1–21. 10.1152/physrev.00017.2012 PubMed DOI

Pasarica M, Sereda OR, Redman LM, Albarado DC, Hymel DT, Roan LE, et al. Reduced adipose tissue oxygenation in human obesity: evidence for rarefaction, macrophage chemotaxis, and inflammation without an angiogenic response. Diabetes. Pennington Biomedical Research Center, Baton Rouge, Louisiana, USA; 2009;58: 718–725. 10.2337/db08-1098 PubMed DOI PMC

Kabon B, Nagele A, Reddy D, Eagon C, Fleshman JW, Sessler DI, et al. Obesity decreases perioperative tissue oxygenation. Anesthesiology. 2004. 10.1097/01.sa.0000144258.38981.73 PubMed DOI PMC

Ye J, Gao Z, Yin J, He Q. Hypoxia is a potential risk factor for chronic inflammation and adiponectin reduction in adipose tissue of ob/ob and dietary obese mice. AmJPhysiol EndocrinolMetab. Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA 70808, USA. yej@pbrc.edu; 2007;293: E1118–E1128. PubMed

Trayhurn P, Wang B, Wood IS. Hypoxia in adipose tissue: a basis for the dysregulation of tissue function in obesity? BrJNutr. 2008; PubMed

Polak J, Studer-Rabeler K, McHugh H, Hussain MA, Shimoda LA. System for exposing cultured cells to intermittent hypoxia utilizing gas permeable cultureware. Gen Physiol Biophys. 2015;34: 235–47. 10.4149/gpb_2014043 PubMed DOI PMC

Ramirez-Zacarias JL, Castro-Munozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry. Department of Cell Biology, Centro de Investigacion y de Estudios Avanzados del IPN, Mexico City, Mexico; 1992;97: 493–497. PubMed

Nunnari JJ, Zand T, Joris I, Majno G. Quantitation of oil red O staining of the aorta in hypercholesterolemic rats. ExpMolPathol. Department of Pathology, University of Massachusetts Medical School, Worcester 01655; 1989;51: 1–8. PubMed

Dyballa N, Metzger S. Fast and sensitive colloidal coomassie G-250 staining for proteins in polyacrylamide gels. JVisExp. Biological Medical Research Center (BMFZ), University of Duesseldorf. Nadine.dyballa@uni-duesseldorf.de; 2009; PubMed PMC

Ye J, Gao Z, Yin J, He Q. Hypoxia is a potential risk factor for chronic inflammation and adiponectin reduction in adipose tissue of ob/ob and dietary obese mice. AmJPhysiol EndocrinolMetab. 2007; PubMed

Pasarica M, Sereda OR, Redman LM, Albarado DC, Hymel DT, Roan LE, et al. Reduced Adipose Tissue Oxygenation in Human Obesity: Evidence for Rarefaction, Macrophage Chemotaxis, and Inflammation Without an Angiogenic Response. Diabetes. 2009;58: 718–725. 10.2337/db08-1098 PubMed DOI PMC

Reinke C, Bevans-Fonti S, Drager LF, Shin MK, Polotsky VY. Effects of different acute hypoxic regimens on tissue oxygen profiles and metabolic outcomes. JApplPhysiol. Dept. of Internal Medicine, Division of Pulmonary Diseases, Univ. Hospital Giessen and Marburg GmbH, 35043 Marburg, Germany. reinke@med.uni-marburg.de; 2011;111: 881–890. PubMed PMC

Wise DR, Ward PS, Shay JE, Cross JR, Gruber JJ, Sachdeva UM, et al. Hypoxia promotes isocitrate dehydrogenase-dependent carboxylation of alpha-ketoglutarate to citrate to support cell growth and viability. ProcNatlAcadSci USA. 2011; PubMed PMC

Des Rosiers C, Fernandez CA, David F, Brunengraber H. Reversibility of the mitochondrial isocitrate dehydrogenase reaction in the perfused rat liver. Evidence from isotopomer analysis of citric acid cycle intermediates. J Biol Chem. 1994;269: 27179–82. Available: http://www.jbc.org/content/269/44/27179.abstract PubMed

Papasian CJ, Hertlein G. Rapid identification of Escherichia coli with a fluorogenic beta-glucuronidase assay. Diagn Microbiol Infect Dis. 1987;8: 255–8. Available: http://www.ncbi.nlm.nih.gov/pubmed/3329592 PubMed

Filipp F V., Scott DA, Ronai ZA, Osterman AL, Smith JW. Reverse TCA cycle flux through isocitrate dehydrogenases 1 and 2 is required for lipogenesis in hypoxic melanoma cells. Pigment Cell Melanoma Res. 2012;25: 375–383. 10.1111/j.1755-148X.2012.00989.x PubMed DOI PMC

Spencer M, Unal R, Zhu B, Rasouli N, McGehee RE, Peterson CA, et al. Adipose tissue extracellular matrix and vascular abnormalities in obesity and insulin resistance. J Clin Endocrinol Metab. 2011;96: E1990–8. 10.1210/jc.2011-1567 PubMed DOI PMC

He Q, Gao Z, Yin J, Zhang J, Yun Z, Ye J. Regulation of HIF-1 activity in adipose tissue by obesity-associated factors: adipogenesis, insulin, and hypoxia. AJP Endocrinol Metab. 2011;300: E877–E885. 10.1152/ajpendo.00626.2010 PubMed DOI PMC

Jiang BH, Semenza GL, Bauer C, Marti HH. Hypoxia-inducible factor 1 levels vary exponentially over a physiologically relevant range of O2 tension. Am J Physiol. 1996;271: C1172–80. Available: http://www.ncbi.nlm.nih.gov/pubmed/8897823 PubMed

Gaglio D, Metallo CM, Gameiro P a, Hiller K, Danna LS, Balestrieri C, et al. Oncogenic K-Ras decouples glucose and glutamine metabolism to support cancer cell growth. Mol Syst Biol. 2011;7: 523 10.1038/msb.2011.56 PubMed DOI PMC

Yoo H, Antoniewicz MR, Stephanopoulos G, Kelleher JK. Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line. J Biol Chem. 2008;283: 20621–7. 10.1074/jbc.M706494200 PubMed DOI PMC

Coppack SW, Persson M, Judd RL, Miles JM. Glycerol and nonesterified fatty acid metabolism in human muscle and adipose tissue in vivo. Am J Physiol. 1999;276: E233–40. Available: http://ajpendo.physiology.org/content/ajpendo/276/2/E233.full.pdf PubMed

MacDougald OA, Burant CF. Obesity and metabolic perturbations after loss of aquaporin 7, the adipose glycerol transporter. Proc Natl Acad Sci U S A. 2005;102: 10759–60. Available: http://www.pnas.org/cgi/content/long/102/31/10759 PubMed PMC

Mráček T, Drahota Z, Houštěk J. The function and the role of the mitochondrial glycerol-3-phosphate dehydrogenase in mammalian tissues. Biochim Biophys Acta. 2013;1827: 401–10. 10.1016/j.bbabio.2012.11.014 PubMed DOI

Gerald D, Berra E, Frapart YM, Chan DA, Giaccia AJ, Mansuy D, et al. JunD reduces tumor angiogenesis by protecting cells from oxidative stress. Cell. Elsevier; 2004;118: 781–94. Available: http://www.cell.com/article/S0092867404008049/fulltext PubMed

Metallo CM, Gameiro PA, Bell EL, Mattaini KR, Yang J, Hiller K, et al. Reductive glutamine metabolism by IDH1 mediates lipogenesis under hypoxia. Nature. Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.; 2012;481: 380–384. Available: 10.1038/nature10602 PubMed DOI PMC

Pavlikova N, Weiszenstein M, Pala J, Halada P, Seda O, Elkalaf M, et al. The Effect Of Cultureware Surfaces On Functional And Structural Components Of Differentiated 3t3-L1 Preadipocytes. Cell Mol Biol Lett. 2015 PubMed

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