Hostname: page-component-7dd5485656-dk7s8 Total loading time: 0 Render date: 2025-10-31T16:24:53.840Z Has data issue: false hasContentIssue false
  • English
  • Français

Short-term modulation of lipogenesis by macronutrients in rainbow trout (Oncorhynchus mykiss) hepatocytes

Published online by Cambridge University Press:  09 March 2007

M. J. Alvarez ,
A. Díez ,
C. López-Bote ,
M. Gallego  and
J. M. Bautista
M. J. Alvarez
Affiliation:
Departamento de Bioquímica y Biología Molecular IV, Universidad Complutense de Madrid, Facultad de Veterinaria, 28040 Madrid, Spain
A. Díez
Affiliation:
Departamento de Bioquímica y Biología Molecular IV, Universidad Complutense de Madrid, Facultad de Veterinaria, 28040 Madrid, Spain
C. López-Bote
Affiliation:
Departamento de Producción Animal, Universidad Complutense de Madrid, Facultad de Veterinaria, 28040 Madrid, Spain
M. Gallego
Affiliation:
Departamento de Bioquímica y Biología Molecular IV, Universidad Complutense de Madrid, Facultad de Veterinaria, 28040 Madrid, Spain
J. M. Bautista*
Affiliation:
Departamento de Bioquímica y Biología Molecular IV, Universidad Complutense de Madrid, Facultad de Veterinaria, 28040 Madrid, Spain
*
*Corresponding author: Dr José M. Bautista, fax +34 91 3943824, email bauchem@eucmax.sim.ucm.es
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Rainbow trout (Oncorhynchus mykiss) hepatocytes were cultured under simulated conditions of varying nutritional status to explore the short-term modulation by dietary substrates of the main lipogenic enzymes: glucose-6-phosphate dehydrogenase (G6PD), malic enzyme (ME), ATP-citrate lyase (ACL), acetyl-CoA carboxylase (ACoAC) and fatty acid synthetase (FAS). Primary cultures were individually exposed to varying amounts of glucose, hydrolysed casein and long-chain polyunsaturated fatty acids (PUFA) for 12 h. A second set of experiments was designed to evaluate the effects of mixing different relative amounts of these macronutrients in the culture medium. Glucose concentrations of up to 20–25 mM SHOWED A STIMULATORY EFFECT ON G6PD, ME, ACL AND ACOAC ACTIVITY (P<0·05) WHILE AN EARLIER INHIBITORY EFFECT ON FAS WAS OBSERVED AT 10–20 Mm glucose (P<0·05). The use of hydrolysed casein as a nutritional source of amino acids inhibited the activity of FAS and ME (P<0·05), and stimulated G6PD, ACoAC and ACL activity (P<0·05). Low levels of linolenic acid exerted a stimulatory effect on all the lipogenic enzymes assayed (P<0·05) with the exception of FAS, and increased amounts showed some inhibition of lipogenic activities (P<0·05). Eicosapentaenoic acid and docosahexaenoic acid showed a similar effect, although the former strongly inhibited FAS activity while the latter showed greater potential to inhibit ACoAC and G6PD. A complete change in the relative levels of glucose, hydrolysed casein and PUFA in turn led to changes in the enzyme activity patterns observed. The present study shows the feasibility of exploring the direct regulation of lipogenesis in isolated fish cells by varying the relative amounts of main macronutrients, mimicking in vivo dietary conditions. It is felt that such an approach may serve to investigate the macronutrient regulation of other metabolic pathways.

Keywords

LipogenesisHepatocytesRainbow trout

Information

Type
Research Article
Information
British Journal of Nutrition , Volume 84 , Issue 5 , November 2000 , pp. 619 - 628
Copyright
Copyright © The Nutrition Society 2000

References

Alvarez, MJ, López-Bote, CJ, Diez, A, Corraze, G, Arzel, J, Dias, J, Kaushik, SJ and Bautista, JM (1998) Dietary fish oil and digestible protein modify susceptibility to lipid peroxidation in the muscle of rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax). British Journal of Nutrition 80, 281–289.CrossRefGoogle ScholarPubMed
Ashida, H, Nakai, R, Kanazawa, K and Danno, G (1998) Xenobiotic tolerance of primary cultured hepatocytes in rats fed a high-fat or high-protein diet. Journal of Nutritional Science and Vitaminology 44, 89–102.Google Scholar
Bautista, JM, Garrido-Pertierra, A and Soler, G (1988) Glucose-6-phosphate dehydrogenase from. Dicentrarchus labrax liver: kinetic mechanism and kinetics of NADPH inhibition. Biochimica et Biophysica Acta 967, 354–363.CrossRefGoogle Scholar
Bell, MV, Henderson, JR and Sargent, JR (1986) The role of polyunsaturated fatty acids in fish. Comparative Biochemistry and Physiology 83B, 711–719.Google Scholar
Bergot, F (1979) Effects of dietary carbohydrates and of their mode of distribution on glycaemia in rainbow trout (Salmo gairdneri R.). Comparative Biochemistry and Physiology 64A, 543–547.CrossRefGoogle Scholar
Böhm, R, Hanke, W and Segner, H (1994) The sequential restoration of plasma metabolite levels, liver composition and liver structure in refed carp,. Cyprinus carpio. Journal of Comparative Physiology 164B, 32–41.CrossRefGoogle Scholar
Bols, NC and Lee, LEJ (1991) Technology and uses of cell cultures from tissues and organs of bony fish. Cytotechnology 6, 163–187.CrossRefGoogle ScholarPubMed
Brauge, C, Medale, F and Corraze, G (1994) Effect of diet carbohydrate levels on growth, body composition and glycaemia in rainbow trout,. Oncorhynchus mykiss, reared in seawater. Aquaculture 123, 109–120.Google Scholar
Chen, RF (1967) Removal of fatty acids from serum albumin by charcoal treatment. Journal of Biological Chemistry 242, 173–181.CrossRefGoogle ScholarPubMed
Clarke, SD and Jump, DB (1994) Dietary polyunsaturated fatty acid regulation of gene transcription. Annual Review of Nutrition 14, 83–98.CrossRefGoogle ScholarPubMed
Dias, J, Alvarez, MJ, Diez, A, Arzel, J, Corraze, G, Bautista, JM and Kaushik, SJ (1998) Regulation of hepatic lipogenesis by dietary protein/energy in juvenile European sea bass (Dicentrarchus labrax). Aquaculture 161, 169–186.CrossRefGoogle Scholar
Foufelle, F, Girard, J and Ferre, P (1996) Regulation of lipogenic enzyme expression by glucose in liver and adipose tissue: a review of the potential cellular and molecular mechanisms. Advances in Enzyme Regulation 36, 199–226.CrossRefGoogle ScholarPubMed
French, CJ, Mommsen, TP and Hochachka, PW (1981) Amino acid utilisation in isolated hepatocytes from rainbow trout. European Journal of Biochemistry 113, 311–317.CrossRefGoogle ScholarPubMed
Fynn-Aikins, K, Hung, SSO, Liu, W and Li, H (1992) Growth, lipogenesis and liver composition of juvenile white sturgeon fed different levels of D-glucose. Aquaculture 105, 61–72.CrossRefGoogle Scholar
Gandemer, G, Durand, G and Pascal, G (1983) Relative contributions of the main tissues and organs to body fatty acid synthesis in the rat. Lipids 18, 223–228.Google Scholar
Geelen, MJH (1994) Medium-chain fatty acids as short-term regulators of hepatic lipogenesis. Biochemical Journal 302, 141–146.CrossRefGoogle ScholarPubMed
Gongnet, GP, Meyerburgdorff, KH, Becker, K and Gunther, KD (1996) Influence of various protein sources on protein digestion and metabolism of growing mirror carp (Cyprinus carpio, L). Revue de Medecine Veterinaire 147, 63–68.Google Scholar
Harmon, JS and Sheridan, MA (1992) Glucose-stimulated lipolysis in rainbow trout. Oncorhynchus mykiss, liver. Fish Physiology and Biochemistry 10, 189–199.CrossRefGoogle ScholarPubMed
Harmon, JS and Sheridan, MA (1992) Effects of nutritional state, insulin and glucagon on lipid mobilization in rainbow trout,. Oncorhynchus mykiss. General and Comparative Endocrinology 87, 214–221.CrossRefGoogle ScholarPubMed
Henderson, RJ (1996) Fatty acid metabolism in freshwater fish with particular reference to polyunsaturated fatty acids. Archives of Animal Nutrition – Archiv fur Tierernahrung 49, 5–22.Google ScholarPubMed
Henderson, RJ, Tocher, DR, (1987) The lipid composition and biochemistry of freshwater fish. In Progress in Lipid Research PP. 281–347.Google Scholar
Hillgartner, FB, Salati, LM and Goodridge, AG (1995) Physiological and molecular mechanisms involved in nutritional regulation of fatty acid synthesis. Physiological Reviews 75, 47–76.Google Scholar
Hilton, JW and Atkinson, JL (1982) Response of rainbow trout (Salmo gairdneri) to increased levels of available carbohydrate in practical trout diets. British Journal of Nutrition 47, 597–607.Google Scholar
Holland, R, Witters, LA and Hardie, DG (1984) Glucagon inhibits fatty acid synthesis in isolated hepatocytes via phosphorylation of acetyl-CoA carboxylase by cyclic AMP-dependent protein kinase. European Journal of Biochemistry 140, 325–333.Google Scholar
Hsu, RY, Butterworth, PHW, Porter, JW (1969) Pigeon liver fatty acid synthetase. In Methods in Enzymology, vol. 14, PP. 33–39 [Lowenstein, JM, editors]. New York, NY: Academic Press Inc.Google Scholar
Hung, SSO and Storebakken, T (1994) Carbohydrate utilisation by rainbow trout is affected by feeding strategy. Journal of Nutrition 124, 223–230.CrossRefGoogle ScholarPubMed
Iritani, N, Ikeda, Y, Fukuda, H and Katsurada, A (1984) Comparative study of lipogenic enzymes in several vertebrates. Lipids 19, 828–835.CrossRefGoogle ScholarPubMed
Klauning, JE (1984) Establishment of fish hepatocyte cultures for use in. in vitro carcinogenicity studies. National Cancer Institute Monographs 65, 163–173.Google Scholar
Klauning, JE, Ruch, RJ and Goldblatt, PJ (1985) Trout hepatocyte culture: isolation and primary culture. In Vitro Cellular and Developmental Biology 21, 221–228.Google Scholar
Leger, C, Fremont, L and Boudon, M (1981) Fatty acid composition of lipids in the trout. Influence of dietary fatty acids on the triglyceride fatty acid desaturation in serum, adipose tissue, liver, white and red muscle. Comparative Biochemistry and Physiology 69B, 99–105.Google Scholar
Leibovitz, A (1963) The growth and maintenance of tissue cell cultures in free gas exchange with the atmosphere. American Journal of Hygiene 78, 173–180.Google ScholarPubMed
Likimani, TA and Wilson, RP (1982) Effects of diet on lipogenic enzyme activities in channel catfish hepatic and adipose tissue. Journal of Nutrition 112, 112–117.CrossRefGoogle ScholarPubMed
Lin, H, Romsos, DR, Tack, PI and Leveille, GAJ (1977) Influence of diet on. in vitro and. in vivo rates of fatty acid synthesis in coho salmon. Journal of Nutrition 107, 1677–1682.Google ScholarPubMed
Moon, TW, Walsh, PJ and Mommsen, TP (1985) Fish hepatocytes. A model metabolic system. Canadian Journal of Fisheries and Aquatic Sciences 42, 1772–1782.CrossRefGoogle Scholar
Newsholme, EA, Start, C, (1973) Regulation of carbohydrate metabolism in liver and regulation of fat metabolism in liver. In Regulation in Metabolism, chapters 6 and 7, pp. 247–316 [Newsholme, EA and Start, C, editors]. New York, NY: John Wiley & Sons.Google Scholar
Ochoa, S, (1955) Malic enzyme. In Methods in Enzymology vol. 1, PP. 739–753.CrossRefGoogle Scholar
Officer, DI, Batterham, ES and Farrel, DJ (1997) Comparison of growth performance and nutrient retention of weaner pigs given diets based on casein, free amino acids or conventional proteins. British Journal of Nutrition 77, 731–744.Google Scholar
Pereira, C, Vijayan, MM and Moon, TW (1995) In-vitro hepatocyte metabolism of alanine and glucose and the response to insulin in fed and fasted rainbow trout. Journal of Experimental Zoology 271, 425–431.CrossRefGoogle Scholar
Segner, H (1998) Isolation and primary culture of teleost hepatocytes. Comparative Biochemistry and Physiology 120A, 71–81.CrossRefGoogle Scholar
Segner, H, Blair, JB, Wirtz, G and Miller, MR (1994) Cultured trout liver cells: utilisation of substrates and response to hormones. In Vitro Cellular and Developmental Biology 30A, 306–311.CrossRefGoogle ScholarPubMed
Segner, H, Böhm, R, (1994) Enzymes of lipogenesis. In Biochemistry and Molecular Biology of Fishes, vol. 3, PP. 313–325.Google Scholar
Sellner, PA and Hazel, JR (1982) Incorporation of polyunsaturated fatty acids into lipids of rainbow trout hepatocytes. American Journal of Physiology 243, R223–R228.Google ScholarPubMed
Sheridan, MA (1994) Regulation of lipid metabolism in poikilothermic vertebrates. Comparative Biochemistry and Physiology 107B, 495–508.Google Scholar
Shikata, T, Masumoto, T and Shimeno, S (1993) Metabolic response to restricted feeding of a high protein diet in common carp. Nippon Suisan Gakkaishi 59, 1587–1591.CrossRefGoogle Scholar
Shimeno, S, Hosokawa, H and Takeda, M (1996) Metabolic response of juvenile yellowtail to dietary carbohydrate to lipid ratios. Fisheries Science 62, 945–949.Google Scholar
Shimeno, S, Kheyyali, D and Shikata, T (1995) Metabolic response to dietary lipid to protein ratios in common carp. Fisheries Science 61, 977–980.CrossRefGoogle Scholar
Simopoulos, AP (1996) The role of fatty acids in gene expression: health implications. Annuals of Nutrition and Metabolism 40, 303–311.Google Scholar
Spector, AA, Mathur, SN, Kaduce, TL and Hyman, BT (1981) Lipid nutrition and metabolism of cultured mammalian cells. Progress of Lipid Research 19, 155–186.CrossRefGoogle Scholar
Srere, PA, (1962) Citrate cleavage enzyme. In Methods in Enzymology, vol. 5, PP. 641–644.Google Scholar
Stabile, LP, Klautky, SA, Minor, SM and Salati, LM (1998) Polyunsaturated fatty acids inhibit the expression of the glucose-6-phosphate dehydrogenase gene in primary rat hepatocytes by a nuclear posttranscriptional mechanism. Journal of Lipid Research 39, 1951–1963.Google Scholar
Suarez, MD, Hidalgo, MC, Garcí Gallego, M, Sanz, A and De la Higuera, M (1995) Influence of the relative proportions of energy yielding nutrients on liver intermediary metabolism of the European eel. Comparative Biochemistry and Physiology 111A, 421–428.Google Scholar
Tocher, DR, Carr, J and Sargent, JR (1989) Polyunsaturated fatty acid metabolism in fish cells: differential metabolism of. n-3 and. n-6 series acids by cultured cells originating from a freshwater teleost fish and from a marine teleost fish. Comparative Biochemistry and Physiology 94B, 367–374.Google Scholar
Tocher, DR, Sargent, JR and Frerichs, GN (1988) The fatty acid compositions of established fish cell lines after long term culture in mammalian sera. Fish Physiology and Biochemistry 5, 219–227.CrossRefGoogle ScholarPubMed
Voss, B and Jankowsky, HD (1986) Temperature-dependence of lipogenesis in isolated hepatocytes from rainbow trout (Salmo gairdneri). Comparative Biochemistry and Physiology B – Biochemistry and Molecular Biology 83, 13–22.CrossRefGoogle ScholarPubMed
Wilson, MD, Blake, WL, Salati, LM and Clarke, SD (1990) Potency of polyunsaturated and saturated fats as short term inhibitors of hepatic lipogenesis in rats. Journal of Nutrition 120, 544–552.Google Scholar
Wilson, RP (1994) Review: utilisation of dietary carbohydrate by fish. Aquaculture 124, 67–80.CrossRefGoogle Scholar
Zampelas, A, Morgan, LM, Furlonger, N and Williams, CM (1995) Effects of dietary fatty acid composition on basal and hormone-stimulated hepatic lipogenesis and on circulating lipids in the rat. British Journal of Nutrition 74, 381–392.Google Scholar