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Effects of fatty acids on skeletal muscle cell differentiation in vitro

Published online by Cambridge University Press:  08 March 2007

Matthew S. Hurley ,
Claire Flux ,
Andrew M. Salter  and
John M. Brameld
Matthew S. Hurley
Affiliation:
University of Nottingham, Division of Nutritional Sciences, School of Biosciences, Sutton Bonington Campus, Loughborough LE12 5RDUK
Claire Flux
Affiliation:
University of Nottingham, Division of Nutritional Sciences, School of Biosciences, Sutton Bonington Campus, Loughborough LE12 5RDUK
Andrew M. Salter
Affiliation:
University of Nottingham, Division of Nutritional Sciences, School of Biosciences, Sutton Bonington Campus, Loughborough LE12 5RDUK
John M. Brameld*
Affiliation:
University of Nottingham, Division of Nutritional Sciences, School of Biosciences, Sutton Bonington Campus, Loughborough LE12 5RDUK
*
*Corresponding author:fax +44 (0)115 951 6122, [email protected]
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Abstract

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Previous studies have shown stimulatory effects of linoleic acid (LA, C18:2) on differentiation of rat muscle cells in culture (Allen et al.1985), but there appears to be little investigation of the effects of other fatty acids. The present study therefore compared the effects of different fatty acids on muscle cell differentiation in vitro. L6 myoblasts were cultured (Dulbecco's Modified Eagles Medium+10% fetal calf serum) in six-well plates until 80% confluent (day 0). Cells were then either harvested or the medium switched to differentiation medium (Dulbecco's Modified Eagles Medium+2% horse serum), supplemented with fatty acid or drug treatments. Cells were harvested on days 0–5 and assayed for creatine kinase (CK), protein and DNA contents, to give a measure of differentiation (CK/DNA). Initial studies indicated a stimulatory effect of the cis9trans11 (c9, t11) isomer of conjugated linoleic acid (CLA) relative to control. By contrast, the trans10, cis12 (t10, c12) isomer of CLA inhibited differentiation. Further experiments indicated that inhibition of differentiation by the t10, c12 CLA isomer was dose-dependent (up to 200μm) and may be via increased cell proliferation. LA and c9, t11 CLA stimulated differentiation at low concentrations (up to 50μm), but inhibited differentiation at high concentrations (200μm). In contrast, oleic acid stimulated differentiation at all concentrations, whereas the saturated fatty acid, palmitic acid, had no effect. The mechanism appeared not to involve either peroxisome proliferator-activated receptors α or γ. The data suggest that only unsaturated fatty acids have an effect and the presence or absence of a cis-9 double bond may be important.

Keywords

Fatty acidsPUFAConjugated linoleic acidSkeletal muscle differentiationMyogenesis
Type
Research Article
Information
British Journal of Nutrition , Volume 95 , Issue 3 , March 2006 , pp. 623 - 630
Copyright
Copyright © The Nutrition Society 2006

References

Allen, RE, Luiten, LS & Dodson, MV 1985 Effect of insulin and linoleic acid on satellite cell differentiation. J Anim Sci 60, 1571–1579.Google Scholar
Belury, MA 2002 Dietary conjugated linoleic acid in health: physiological effects and mechanisms of action. Annu Rev Nutr 22, 505–531.CrossRefGoogle ScholarPubMed
Brameld, JM, Buttery, PJ, Dawson, JM & Harper, JMM 1998 Nutritional and hormonal control of skeletal-muscle cell growth and differentiation. Proc Nutr Soc 57, 207–217.Google Scholar
Brodie, AE, Manning, VA, Ferguson, KR, Jewell, DE & Hu, CY 1999 Conjugated linoleic acid inhibits differentiation of pre- and postconfluent 3T3-L1 preadipocytes but inhibits cell proliferation only in preconfluent cells. J Nutr 129, 602–606.Google Scholar
Brown, JM, Halvorsen, YD, Lea-Currie, YR, Geigerman, C & McIntosh, M 2001 Trans-10, cis-12, but not cis-9, trans-11, conjugated linoleic acid attenuates lipogenesis in primary cultures of stromal vascular cells from human adipose tissue. J Nutr 131, 2316–2321.Google Scholar
Brun, RP, Kim, JB, Hu, E, Altiok, S & Spiegelman, BM 1996 Adipocyte differentiation: a transcriptional regulatory cascade. Curr Opin Cell Biol 8, 826–832.CrossRefGoogle ScholarPubMed
Clarke, SD 2000 Polyunsaturated fatty acid regulation of gene transcription: a mechanism to improve energy balance and insulin resistance. Br J Nutr 83, Suppl. 1, S59–S66.Google Scholar
Clemente, CFMZ, Corat, MAF, Saad, STO & Klebere, G 2005 Differentiation of C2C12 myoblasts is critically regulated by FAK signalling. Am J Physiol 289, R862–R870.Google Scholar
Ding, S-T & Mersmann, HJ 2001 Fatty acids modulate porcine adipocyte differentiation and transcripts for transcription factors and adipocyte-characteristic proteins. J Nutr Biochem 12, 101–108.Google Scholar
Florini, JR, Magri, KA, Ewton, DZ, James, PL, Grindstaff, K & Rotwein, PS 1991 ‘Spontaneous’ differentiation of skeletal myoblasts is dependent upon autocrine secretion of insulin-like growth factor II. J Biol Chem 266, 15917–15923.CrossRefGoogle ScholarPubMed
Goldstein, JL, Basu, SK & Brown, MS 1983 Receptor-mediated endocytosis of low-density lipoprotein in cultured cells. Methods Enzymol 98, 241–260.Google Scholar
Grimaldi, PA 2005 Regulatory role of peroxisome proliferator-activated receptor delta (PPARd) in muscle metabolism. A new target for metabolic syndrome treatment?. Biochimie 87, 5–8.CrossRefGoogle Scholar
Holst, D, Luquet, S, Nogueira, V, Kristiansen, K, Leverve, X & Grimaldi, PA 2003 Nutritional regulation and role of peroxisome proliferator-activated receptor d in fatty acid catabolism in skeletal muscle. Biochim Biophys Acta 1633, 43–50.CrossRefGoogle Scholar
Kelly, GS 2001 Conjugated linoleic acid: a review. Altern Med Rev 6, 367–382.Google Scholar
Leung, YH & Liu, RH 2000 trans-10, cis-12-Conjugated linoleic acid isomer exhibits stronger oxyradical scavenging capacity than cis-9, trans-11-conjugated linoleic acid isomer. J Agric Food Chem 48, 5469–5475.Google Scholar
Lin, Y, Kreeft, A, Schuurbiers, JAE & Draijer, R 2001 Different effects of conjugated linoleic acid isomers on lipoprotein lipase activity in 3T3-L1 adipocytes. J Nutr Biochem 12, 183–189.Google Scholar
Lowry, OH, Rosebrough, NJ, Farr, AL & Randall, RJ 1951 Protein measurement with the Folin phenol reagent. J Biol Chem 193, 265–275.CrossRefGoogle ScholarPubMed
Mitsiades, CS, Mitsiades, N, Poulaki, V et al. , 2002 Activation of NFkappaB and upregulation of intracellular anti-apoptotic proteins via the IGF-1/Akt signaling in human multiple myeloma cells: therapeutic implications. Oncogene 21, 5673–5683.Google Scholar
Orzechowski, A, Grizard, J, Jank, M, Gajkowska, B, Lokociejewska, M, Zaron-Teperek, M & Godlewski, M 2002 Dexamethasonemediated regulation of death and differentiation of muscle cells. Is hydrogen peroxide involved in the process?. Reprod Nutr Dev 42, 197–216.CrossRefGoogle ScholarPubMed
Pariza, MW, Park, Y & Cook, ME 2001 The biologically active isomers of conjugated linoleic acid. Prog Lipid Res 40, 283–298.Google Scholar
Rago, R, Mitchen, J & Wilding, G 1990 DNA fluorometric assay in 96-well tissue-culture plates using Hoechst-33258 after cell-lysis by freezing in distilled water. Anal Biochem. 191, 31–34.Google Scholar
Siebenlist, U, Franzoso, G & Brown, K 1994 Structure, regulation and function of NF-kappaB. Annu Rev Cell Biol 10, 405–455.Google Scholar
Van Harken, DR, Dixon, CW & Heimberg, M 1969 Hepatic lipid metabolism in experimental diabetes V. The effect of concentration of oleate on metabolism of triglycerides and on ketogenesis. J Biol Chem 244, 2278–2285.Google Scholar
Wolf, G 2004 Tissue-specific knockout defines peroxisome proliferator-activated receptor gamma function in muscle and liver. Nutr Rev 62, 253–255.Google Scholar