Hostname: page-component-586b7cd67f-t7czq Total loading time: 0 Render date: 2024-11-26T11:03:02.877Z Has data issue: false hasContentIssue false
  • English
  • Français

Regulation of lipid accumulation in 3T3-L1 cells: insulin-independent and combined effects of fatty acids and insulin

Published online by Cambridge University Press:  01 January 2008

T. A. Kokta ,
A. L. Strat ,
M. R. Papasani ,
J. I. Szasz ,
M. V. Dodson  and
R. A. Hill
T. A. Kokta
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
A. L. Strat
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
M. R. Papasani
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
J. I. Szasz
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
M. V. Dodson
Affiliation:
Department of Animal Sciences, Washington State University, Pullman, WA 99164, USA
R. A. Hill*
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
*
E-mail: [email protected]
Get access

Abstract

The insulin-independent and combined effects of fatty acids (FA; linoleic and oleic acids) and insulin in modulating lipid accumulation and adipogenesis in 3T3-L1 cells was investigated using a novel protocol avoiding the effects of a complex hormone ‘induction’ mixture. 3T3-L1 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) plus serum (control) or in DMEM plus either 0.3 mmol/l linoleic or oleic acids with 0.3 mmol/l FA-free bovine serum albumin in the presence or absence of insulin. Cells were cultured for 4 to 8 days and cell number, lipid accumulation, peroxisome proliferator-activated receptor-gamma (PPAR-γ) and glucose transporter 4 (GLUT-4) protein expression were determined. Cell number appeared to be decreased in comparison with control cultures. In both oleic acid and linoleic acid-treated cells, notably in the absence (and presence) of insulin, oil-red O stain-positive cells showed abundant lipid. The percentage of cells showing lipid accumulation was greater in FA-treated cultures compared with control cells grown in DMEM plus serum (P < 0.001). Treatment with both linoleic and oleic acid-containing media evoked higher levels of PPAR-γ than observed in control cultures (P < 0.05). GLUT-4 protein also increased in response to treatment with both linoleic and oleic acid-containing media (P < 0.001). Lipid accumulation in 3T3-L1 cells occurs in response to either oleic or linoleic acids independently of the presence of insulin. Both PPAR-γ and GLUT-4 protein expression were stimulated. Both proteins are considered markers of adipogenesis, and these observations suggest that these cells had entered the physiological state broadly accepted as differentiated. Furthermore, 3T3-L1 cells can be induced to accumulate lipid in a serum-free medium supplemented with FA, without the use of induction protocols using complex hormone mixtures. We have demonstrated a novel model for the study of lipid accumulation that will improve the understanding of adipogenesis in adipocyte lineage cells.

Keywords

adipogenesisfatty acidsGLUT-4insulinPPAR-γ
Type
Full Paper
Information
animal , Volume 2 , Issue 1 , January 2008 , pp. 92 - 99
Copyright
Copyright © The Animal Consortium 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Amri, EZ, Bonino, F, Ailhaud, G, Abumrad, NA, Grimaldi, PA 1995. Cloning of a protein that mediates transcriptional effects of fatty acids in preadipocytes. Homology to peroxisome proliferator-activated receptors. Journal of Biological Chemistry 270, 2367–2371.CrossRefGoogle ScholarPubMed
Bishop-Bailey, D, Wray, J 2003. Peroxisome proliferator-activated receptors: a critical review on endogenous pathways for ligand generation. Prostaglandins and Other Lipid Mediators 71, 1–22.CrossRefGoogle ScholarPubMed
Boone, C, Mourot, J, Gregiore, F, Remacle, C 2000. The adipose conversion process: regulation by extracellular and intracellular factors. Reproduction, Nutrition, Development 40, 325–358.CrossRefGoogle ScholarPubMed
Brodie, AE, Manning, VA, Ferguson, KR, Jewell, DE, Hu, CY 1999. Conjugated linoleic acid inhibits differentiation of pre- and post-confluent 3T3-L1 preadipocytes but inhibits cell proliferation only in preconfluent cells. Journal of Nutrition 129, 602–606.CrossRefGoogle ScholarPubMed
Brown, JM, McIntosh, MK 2003. Conjugated linoleic acid in humans: regulation of adiposity and insulin sensitivity. Journal of Nutrition 133, 3041–3046.CrossRefGoogle ScholarPubMed
Chiaramonte, R, Bartolini, E, Testolin, C, Comi, P 1998. Regulation of the human GLUT4 gene expression in tumor RD18 cell line. Pathobiology 66, 191–195.CrossRefGoogle ScholarPubMed
Croissandeau, G, Chretien, M, Mbikay, M 2002. Involvement of matrix metalloproteinases in the adipose conversion of 3T3-L1 preadipocytes. Biochemical Journal 364, 739–746.CrossRefGoogle ScholarPubMed
De Vries, J, Vork, M, Roemen, T, De Jong, Y, Cleutjens, J, Van der Vusse, G, Van Bilsen, M 1997. Saturated but not mono-unsaturated fatty acids induce apoptotic cell death in neonatal rat ventricular myocytes. Journal of Lipid Research 38, 1384–1394.CrossRefGoogle Scholar
Ding, ST, Mersmann, HJ 2001. Fatty acids modulate porcine adipocyte differentiation and transcripts for transcription factors and adipocyte-characteristic proteins. Journal of Nutritional Biochemistry 12, 101–108.Google ScholarPubMed
Ding, ST, McNeel, RL, Mersmann, HJ 2002. Modulation of adipocyte determination and differentiation-dependent factor 1 by selected polyunsaturated fatty acids. In vitro Cellular and Developmental Biology – Animal 38, 352–357.2.0.CO;2>CrossRefGoogle ScholarPubMed
El-Jack, AK, Kandror, KV, Pilch, PF 1999. The formation of an insulin-responsive vesicular cargo compartment is an early event in 3T3-L1 adipocyte differentiation. Molecular Biology of the Cell 10, 1581–1594.CrossRefGoogle ScholarPubMed
Ferré, P 2004. The biology of peroxisome proliferator-activated receptors: relationship with lipid metabolism and insulin sensitivity. Diabetes 53 (Suppl. 1), S43–S50.CrossRefGoogle ScholarPubMed
Floyd, ZE, Stephens, JM 2003. STAT5A promotes adipogenesis in nonprecursor cells and associates with the glucocorticoid receptor during adipocyte differentiation. Diabetes 52, 308–314.CrossRefGoogle ScholarPubMed
Forman, BM, Chen, J, Evans, RM 1997. Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors alpha and delta. Proceedings of the National Academy of Sciences of the United States of America 94, 4312–4317.CrossRefGoogle ScholarPubMed
Grimaldi, PA 2001. The roles of PPARs in adipocyte differentiation. Progress in Lipid Research 40, 269–281.CrossRefGoogle ScholarPubMed
Gross, DN, Farmer, SR, Pilch, PF 2004. Glut4 storage vesicles without Glut4: transcriptional regulation of insulin-dependent vesicular traffic. Molecular and Cellular Biology 24, 7151–7162.CrossRefGoogle ScholarPubMed
Guo, W, Choi, JK, Kirkland, JL, Corkey, BE, Hamilton, JA 2000. Esterification of free fatty acids in adipocytes: a comparison between octanoate and oleate. Biochemical Journal 349, 463–471.CrossRefGoogle ScholarPubMed
Habinowski, SA, Witters, LA 2001. The effects of AICAR on adipocyte differentiation of 3T3-L1 cells. Biochemical and Biophysical Research Communications 286, 852–856.CrossRefGoogle ScholarPubMed
Han, J, Farmer, SR, Kirkland, JL, Corkey, BE, Yoon, R, Pirtskhalava, T, Ido, Y, Guo, W 2002. Octanoate attenuates adipogenesis in 3T3-L1 preadipocytes. Journal of Nutrition 132, 904–910.CrossRefGoogle ScholarPubMed
Hill, RA, Strat, AL, Hughes, NJ, Kokta, TJ, Dodson, MV, Gertler, A 2004. Early insulin signaling cascade in a model of oxidative skeletal muscle: mouse Sol8 cell line. Biochimica et Biophysica Acta 1693, 205–211.CrossRefGoogle Scholar
Hsu, JM, Ding, ST 2003. Effect of polyunsaturated fatty acids on the expression of transcription factor adipocyte determination and differentiation-dependent factor 1 and of lipogenic and fatty acid oxidation enzymes in porcine differentiating adipocytes. British Journal of Nutrition 90, 507–513.CrossRefGoogle ScholarPubMed
Hutley, LJ, Newell, FM, Joyner, JM, Suchting, SJ, Herington, AC, Cameron, DP, Prins, JB 2003. Effects of rosiglitazone and linoleic acid on human preadipocyte differentiation. European Journal of Clinical Investigation 33, 574–581.CrossRefGoogle ScholarPubMed
Imai, T, Takakuwa, R, Marchand, S, Dentz, E, Bornert, JM, Messaddeq, N, Wendling, O, Mark, M, Desvergne, B, Wahli, W, Chambon, P, Metzger, D 2004. Peroxisome proliferator-activated receptor gamma is required in mature white and brown adipocytes for their survival in the mouse. Proceedings of the National Academy of Sciences of the United States of America 101, 4543–4547.CrossRefGoogle Scholar
Kang, K, Pariza, MW 2001. trans-10, cis-12-conjugated linoleic acid reduces leptin secretion from 3T3-L1 adipocytes. Biochemical and Biophysical Research Communications 287, 377–382.CrossRefGoogle ScholarPubMed
Kang, K, Liu, W, Albright, KJ, Park, Y, Pariza, MW 2003. trans-10, cis-12 CLA inhibits differentiation of 3T3-L1 adipocytes and decreases PPAR gamma expression. Biochemical and Biophysical Research Communications 303, 795–799.CrossRefGoogle ScholarPubMed
Kersten, S, Mandard, S, Tan, NS, Escher, P, Metzger, D, Chambon, P, Gonzalez, FJ, Desvergne, B, Wahli, W 2000. Characterization of the fasting-induced adipose factor FIAF, a novel peroxisome proliferator-activated receptor target gene. Journal of Biological Chemistry 275, 28488–28493.CrossRefGoogle ScholarPubMed
Kokta, TA, Dodson, MV, Gertler, A, Hill, RA 2004. Intercellular signaling between adipose tissue and muscle tissue. Domestic Animal Endocrinology 27, 303–331.CrossRefGoogle ScholarPubMed
Lacasa, D, Garcia Dos Santos, E, Giudicelli, Y 2001. Site-specific control of rat preadipocyte adipose conversion by ovarian status: possible involvement of CCAAT/enhancer-binding protein transcription factors. Endocrine 15, 103–110.CrossRefGoogle ScholarPubMed
Lee, K, Hausman, DB, Dean, RG 1999. Expression of CCAAT/enhancer binding protein C/EBPα, β and δ in rat adipose stromal-vascular cells in vitro. Biochimica et Biophysica Acta 1450, 397–405.CrossRefGoogle ScholarPubMed
Li, Y, Lazar, MA 2002. Differential gene regulation by PPARgamma agonist and constitutively active PPARgamma2. Molecular Endocrinology 16, 1040–1048.Google ScholarPubMed
Listenberger, LL, Han, X, Lewis, SE, Cases, S, JrFarese, RV, Ory, DS, Schaffer, JE 2003. Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proceedings of the National Academy of Sciences of the United States of America 100, 3077–3082.CrossRefGoogle ScholarPubMed
McNeel, RL, Mersmann, HJ 2003. Effects of isomers of conjugated linoleic acid on porcine adipocyte growth and differentiation. Journal of Nutritional Biochemistry 14, 266–274.CrossRefGoogle ScholarPubMed
McNeel, RL, Smith, EO, Mersmann, HJ 2003. Isomers of conjugated linoleic acid modulate human preadipocyte differentiation. In Vitro Cellular and Developmental Biology – Animal 39, 375–382.2.0.CO;2>CrossRefGoogle ScholarPubMed
Margetic, S, Gazzola, C, Pegg, GG, Hill, RA 2002. Leptin: a review of its peripheral actions and interactions. International Journal of Obesity and Related Metabolic Disorders 26, 1407–1433.CrossRefGoogle Scholar
Sakaue, H, Ogawa, W, Matsumoto, M, Kuroda, S, Takata, M, Sugimoto, T, Spiegelman, BM, Kasuga, M 1998. Posttranscriptional control of adipocyte differentiation through activation of phosphoinositide 3-kinase. Journal of Biological Chemistry 273, 28945–28952.CrossRefGoogle ScholarPubMed
Schneider, N, Lanz, S, Ramer, R, Schaefer, D, GoppeltStruebe, M 2001. Up-regulation of cyclooxygenase-1 in neuroblastoma cell lines by retinoic acid and corticosteroids. Journal of Neurochemistry 77, 416–424.CrossRefGoogle ScholarPubMed
Shin, SM, Kim, K, Kim, JK, Yoon, SR, Choi, I, Yang, Y 2003. Dexamethasone reverses TGF-beta-mediated inhibition of primary rat preadipocyte differentiation. Federation of European Biochemical Societies Letters 543, 25–30.CrossRefGoogle ScholarPubMed
Spiegelman, BM 1998. PPAR-gamma: adipogenic regulator and thiazolidinedione receptor. Diabetes 47, 507–514.CrossRefGoogle ScholarPubMed
Statistical Analysis Systems Institute 2002. SAS version 8.2. SAS Institute Inc., Cary, NC, USA.Google Scholar
Thompson, DL, Lum, KD, Nygaard, SC, Kuestner, RE, Kelly, KA, Gimble, JM, Moore, EE 1998. The derivation and characterization of stromal cell lines from the bone marrow of p53−/− mice: new insights into osteoblast and adipocyte differentiation. Journal of Bone and Mineral Research 13, 195–204.CrossRefGoogle ScholarPubMed
Tontonoz, P, Hu, E, Spiegelman, BM 1994a. Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell 79, 1147–1156.CrossRefGoogle ScholarPubMed
Tontonoz, P, Hu, E, Graves, RA, Budavari, AI, Spiegelman, BM 1994b. mPPAR gamma 2: tissue-specific regulator of an adipocyte enhancer. Genes and Development 8, 1224–1234.CrossRefGoogle ScholarPubMed
Torii, SI, Kawada, T, Matsuda, K, Matsui, T, Ishihara, T, Yano, H 1998. Thiazolidinedione induces the adipose differentiation of fibroblast-like cells resident within bovine skeletal muscle. Cell Biology International 22, 421–427.CrossRefGoogle ScholarPubMed
Wu, Z, Bucher, NL, Farmer, SR 1996. Induction of peroxisome proliferator-activated receptor gamma during the conversion of 3T3 fibroblasts into adipocytes is mediated by C/EBPbeta, C/EBPdelta, and glucocorticoids. Molecular and Cellular Biology 16, 4128–4136.CrossRefGoogle Scholar
Wu, Z, Xie, Y, Morrison, RF, Bucher, NL, Farmer, SR 1998. PPARgamma induces the insulin-dependent glucose transporter GLUT4 in the absence of C/EBPalpha during the conversion of 3T3 fibroblasts into adipocytes. Journal of Clinical Investigation 101, 22–32.CrossRefGoogle ScholarPubMed
Yamauchi, T, Kamon, J, Waki, H, Murakami, K, Motojima, K, Komeda, K, Ide, T, Kubota, N, Terauchi, Y, Tobe, K, Miki, H, Tsuchida, A, Akanuma, Y, Nagai, R, Kimura, S, Kadowaki, T 2001. The mechanisms by which both heterozygous peroxisome proliferator-activated receptor gamma (PPARgamma) deficiency and PPARgamma agonist improve insulin resistance. Journal of Biological Chemistry 276, 41245–41254.CrossRefGoogle ScholarPubMed