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Metabolic regulation of fatty acid esterification and effects of conjugated linoleic acid on glucose homeostasis in pig hepatocytes

Published online by Cambridge University Press:  22 September 2011

J. A. Conde-Aguilera
Affiliation:
Departamento de Fisiología y Bioquímica de la Nutrición Animal, Estación Experimental del Zaidín (INA CSIC), Profesor Albareda 1, 18008 Granada, Spain
M. Lachica
Affiliation:
Departamento de Fisiología y Bioquímica de la Nutrición Animal, Estación Experimental del Zaidín (INA CSIC), Profesor Albareda 1, 18008 Granada, Spain
R. Nieto
Affiliation:
Departamento de Fisiología y Bioquímica de la Nutrición Animal, Estación Experimental del Zaidín (INA CSIC), Profesor Albareda 1, 18008 Granada, Spain
I. Fernández-Fígares*
Affiliation:
Departamento de Fisiología y Bioquímica de la Nutrición Animal, Estación Experimental del Zaidín (INA CSIC), Profesor Albareda 1, 18008 Granada, Spain
*
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Abstract

Conjugated linoleic acids (CLAs) are geometric and positional isomers of linoleic acid (LA) that promote growth, alter glucose metabolism and decrease body fat in growing animals, although the mechanisms are poorly understood. A study was conducted to elucidate the effects of CLA on glucose metabolism, triglyceride (TG) synthesis and IGF-1 synthesis in primary culture of porcine hepatocytes. In addition, hormonal regulation of TG and IGF-1 synthesis was addressed. Hepatocytes were isolated from piglets (n = 5, 16.0 ± 1.98 kg average body weight) by collagenase perfusion and seeded into collagen-coated T-25 flasks. Hepatocytes were cultured in William's E containing dexamethasone (10−8 and 10−7 M), insulin (10 and 100 ng/ml), glucagon (0 and 100 ng/ml) and CLA (1 : 1 mixture of cis-9, trans-11 and trans-10, cis-12 CLA, 0.05 and 0.10 mM) or LA (0.05 and 0.10 mM). Addition of CLA decreased gluconeogenesis (P < 0.05), whereas glycogen synthesis and degradation, TG synthesis and IGF-1 synthesis were not affected compared with LA. Increased concentration of fatty acids in the media decreased IGF-1 production (P < 0.001) and glycogen synthesis (P < 0.01), and increased gluconeogenesis (P < 0.001) and TG synthesis (P < 0.001). IGF-1 synthesis increased (P < 0.001) and TG synthesis decreased (P < 0.001) as dexamethasone concentration in the media rose. High insulin/glucagon increased TG synthesis. These results indicate that TG synthesis in porcine hepatocytes is hormonally regulated so that dexamethasone decreases and insulin/glucagon increases it. In addition, CLA decreases hepatic glucose production through decreased gluconeogenesis.

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Full Paper
Copyright
Copyright © The Animal Consortium 2011

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References

Agius, L, Chowdhury, MH, Alberti, KGMM 1986. Regulation of ketogenesis, gluconeogenesis and the mitochondrial redox state by dexamethasone in hepatocytem monolayer cultures. Biochemical Journal 239, 593601.CrossRefGoogle Scholar
Ajuwon, KM, Kuske, JL, Ragland, D, Adeola, O, Hancock, DL, Anderson, DB, Spurlock, ME 2003. The regulation of IGF-1 by leptin in the pig is tissue specific and independent of changes in growth hormone. Journal of Nutritional Biochemistry 14, 522530.Google Scholar
Amatruda, JM, Danahy, SA, Chang, CL 1983. The effects of glucocorticoids on insulin-stimulated lipogenesis in primary cultures of rat hepatocytes. Biochemical Journal 212, 135141.CrossRefGoogle ScholarPubMed
Ametaj, BN, Bobe, G, Lu, Y, Young, JW, Beitz, DC 2003. Effect of sample preparation, length of time, and sample size on quantification of total lipids from bovine liver. Journal of Agricultural and Food Chemistry 51, 21052110.Google Scholar
Arany, E, Strain, AJ, Hube, MJ, Phillips, ID, Hill, DJ 1993. Interactive effects of nutrients and hormones on the expression of insulin-like growth factor binding protein-I (IGFBP-I) mRNA and peptide, and IGF-1 release from isolated adult rat hepatocytes. Journal of Cellular Physiology 155, 426435.CrossRefGoogle Scholar
Beauloye, V, Ketelslegers, JM, Moreau, B, Thissen, JP 1999. Dexamethasone inhibits both growth hormone (GH)-induction of insulin-like growth factor-I (IGF-I) mRNA and GH receptor (GHR) mRNA levels in rat primary cultured hepatocytes. Growth Hormone & IGF Research 9, 205211.Google Scholar
Behnia, K, Bhatia, S, Jastromb, N, Balis, U, Sullivan, S, Yarmush, M, Toner, M 2000. Xenobiotic metabolism by cultured primary porcine hepatocytes. Tissue Engineering 6, 467479.CrossRefGoogle ScholarPubMed
Boyd, RD, Whitehead, DM, Butler, WR 1985. Effect of exogenous glucagon and free fatty-acids on gluconeogenesis in fasting neonatal pigs. Journal of Animal Science 60, 659665.Google Scholar
Brameld, JM, Weller, PA, Saunders, JC, Buttery, PJ, Gilmour, RS 1995. Hormonal control of insulin-like growth factor-I and growth hormone receptor mRNA expression by porcine hepatocytes in culture. Journal of Endocrinology 146, 239245.Google Scholar
Calder, PC, Bond, JA, Harvey, DJ, Gordon, S, Newsholme, EA 1990. Uptake and incorporation of saturated and unsaturated fatty acids into macrophage lipids and their effect upon macrophage adhesion and phagocytosis. Biochemistry Journal 269, 807814.Google Scholar
Cantwell, H, Devery, R, Oshea, M, Stanton, C 1999. The effect of conjugated linoleic acid on the antioxidant enzyme defense system in rat hepatocytes. Lipids 34, 833839.CrossRefGoogle ScholarPubMed
Christiansen, RZ 1977. Regulation of palmitate metabolism by carnitine and glucagon in hepatocytes isolated from fasted and carbohydrate refed rats. Biochimica et Biophysica Acta 488, 249262.CrossRefGoogle ScholarPubMed
Clement, L, Poirier, H, Niot, I, Bocher, V, Guerre-Millo, M, Krief, S, Staels, B, Besnard, P 2002. Dietary trans-10,cis-12 conjugated linoleic acid induces hyperinsulinemia and fatty liver in the mouse. Journal of Lipid Research 43, 14001409.CrossRefGoogle ScholarPubMed
Clore, JN, Glickman, PS, Helm, ST, Nestler, JE, Blackard, WG 1991. Evidence for dual control mechanism regulating hepatic glucose output in nondiabetic men. Diabetes 40, 10331040.CrossRefGoogle ScholarPubMed
Corl, BA, Oliver, SAM, Lin, X, Oliver, WT, Ma, Y, Harrell, RJ, Odle, J 2008. Conjugated linoleic acid reduces body fat accretion and lipogenic gene expression in neonatal pigs fed low- or high-fat formulas. Journal of Nutrition 138, 449454.Google Scholar
Diamant, S, Shafrir, E 1975. Modulation of the activity of insulin-dependent enzymes of lipogenesis by glucocorticoids. European Journal of Biochemistry 53, 541546.CrossRefGoogle ScholarPubMed
Duée, PH, Pégorier, JP, Peret, J, Girard, J 1985. Separate effects of fatty acid oxidation and glucagon on gluconeogenesis in isolated hepatocytes from newborn pigs. Biology of the Neonate 47, 7783.Google Scholar
Duerden, JM, Gibbons, GF 1990. Storage, mobilization and secretion of cytosolic triacylglycerol in hepatocyte cultures. The role of insulin. Biochemistry Journal 272, 583587.Google Scholar
Dugan, MER, Aalhus, JL, Schaefer, AL, Kramer, JKG 1997. The effect of conjugated linoleic acid on fat to lean repartitioning and feed conversion in pigs. Canadian Journal of Animal Science 77, 723725.Google Scholar
Dunger, D, Yuen, K, Ong, K 2004. Insulin-like growth factor I and impaired glucose tolerance. Hormone Research 62, 101107.Google Scholar
Fenton, JP, Roehrig, KL, Mahan, DC, Corley, JR 1985. Effect of swine weaning age on body fat and lipogenic activity in liver and adipose tissue. Journal of Animal Science 60, 190199.Google Scholar
Fernández-Fígares, I, Shannon, AE, Wray-Cahen, D, Caperna, TJ 2004. The role of insulin, glucagon, dexamethasone and leptin in the regulation of ketogenesis and glycogen storage in primary cultures of porcine hepatocytes prepared from growing pigs. Domestic Animal Endocrinology 27, 125140.Google Scholar
Fernández-Fígares, I, Lachica, M, Nieto, R, Aguilera, JF 2007. Serum profile of metabolites and hormones of growing Iberian gilts fed diets supplemented with betaine, conjugated linoleic acid or both2nd International Symposium on Energy and Protein Metabolism and Nutrition (ed. I Ortigues-Marty, N Miraux, W Brand-Williams), Vichy, France, pp. 385386. Wageningen Academic Publishers, Wageningen, The Netherlands.CrossRefGoogle Scholar
Ferré, P, Satabin, P, El Manoubi, L, Callikan, S, Girard, J 1981. Relationship between ketogenesis and gluconeogenesis in isolated hepatocytes from newborn rats. Biochemistry Journal 200, 429433.CrossRefGoogle ScholarPubMed
Gondret, F, Ferré, P, Dugail, I 2001. ADD-1/SREBP-1 is a major determinant of tissue differential lipogenic capacity in mammalian and avian species. Journal of Lipid Research 42, 106113.Google Scholar
Granlund, L, Larsen, LN, Nebb, HI, Pedersen, JI 2005. Effects of structural changes of fatty acids on lipid accumulation in adipocytes and primary hepatocytes. Biochimica et Biophysica Acta 1687, 2330.CrossRefGoogle ScholarPubMed
Gregory, PG, Connolly, CK, Toner, M, Sullivan, SJ 2000. In vitro characterization of porcine hepatocyte function. Cell Transplantation 9, 110.Google Scholar
Guillouzo, A 1986. Use of isolated and cultured hepatocytes for xenobiotic metabolism and cytotoxicity studies. In Research in isolated and cultures hepatocytes (ed. A Guillouzo and C Guguen-Guillouzo), pp. 313332. Les Editions INSERM/John Libbey Eurotext Ltd, Paris and London, France and UK.Google Scholar
Hansson, PK, Asztély, A, Clapham, JC, Schreyer, SA 2004. Glucose and fatty acid metabolism in McA-RH7777 hepatoma cells vs. rat primary hepatocytes: responsiveness to nutrient availability. Biochimica et Biophysica Acta 1684, 5462.Google Scholar
Houseknecht, KL, Vanden Heuvel, JP, Moya-Camarena, SY, Portocarrero, CP, Peck, LW, Nickel, KP, Belury, MA 1998. Dietary conjugated linoleic acid normalizes impaired glucose tolerance in the Zucker diabetic fatty fa/fa rat. Biochemical and Biophysical Research Communications 244, 678682.CrossRefGoogle ScholarPubMed
José, AAFBV, Gama, MAS, Lanna, DDP 2008. Effects of trans-10, cis-12 conjugated linoleic acid on gene expression and lipid metabolism of adipose tissue of growing pigs. Genetics and Molecular Research 7, 284294.Google Scholar
Koebe, HG, Schildberg, FW 1996. Isolation of porcine hepatocytes from slaughterhouse organs. International Journal of Artificial Organs 19, 5360.Google Scholar
Lee, KC, Azain, MJ, Hausman, DB, Ramsay, TG 2000. Somatotropin and adipose tissue metabolism: substrate and temporal effects. Journal of Animal Science 78, 12361246.Google Scholar
Lepine, AJ, Boyd, RD, Whitehead, DM 1991. Effect of colostrum intake on hepatic gluconeogenesis and fatty acid oxidation in the neonatal pig. Journal of Animal Science 69, 19661974.Google Scholar
Lepine, AJ, Watford, M, Boyd, RD, Ross, DA, Whitehead, DM 1993. Relationship between hepatic fatty acid oxidation and gluconeogenesis in the fasting neonatal pig. British Journal of Nutrition 70, 8191.CrossRefGoogle ScholarPubMed
Li, Y, Seifert, MF, Ney, DM, Grahn, M, Grant, AL, Allen, KG, Watkins, BA 1999. Dietary conjugated linoleic acids alter serum IGF-1 and IGF binding protein concentrations and reduce bone formation in rats fed (n-6) or (n-3) fatty acids. Journal of Bone and Mineral Research 14, 11531162.CrossRefGoogle ScholarPubMed
Mantha, L, Palacios, E, Deshaies, Y 1999. Modulation of triglyceride metabolism by glucocorticoids in diet-induced obesity. American Journal of Physiology 277, R455R464.Google ScholarPubMed
McGarry, JD, Foster, DW 1980. Effects of exogenous fatty acid concentration on glucagon-induced changes in hepatic fatty acid metabolism. Diabetes 29, 236240.CrossRefGoogle ScholarPubMed
Nelson, DL, Cox, MM 2000. The citric acid cycle. In Lehninger principles of biochemistry, 3rd edition (ed. DL Nelson and MM Cox), pp. 567597. Worth Publishers, New York, NY, USA.Google Scholar
Nerurkar, LS, Marino, PA, Adams, DO 1981. Quantification of selected intracellular and secreted hydrolases of macrophages. In Manual of macrophage methodology(ed. HB Herscowitz, HT Holden, JA Bellanti and A Ghaffer), pp. 229247. Marcel Dekker Inc., New York, NY, USA.Google Scholar
Nordlie, RC, Sukalski, A, Alvares, FL 1980. Responses of glucose 6-phosphate levels to varied glucose loads in the isolated perfused rat liver. Journal of Biological Chemistry 255, 18341838.Google Scholar
Pariza, MW, Park, Y 2001. The biologically active isomers of conjugated linoleic acid. Progress in Lipid Research 40, 283298.CrossRefGoogle ScholarPubMed
Pégorier, JP, Duée, PH, Girard, J, Peret, J 1983. Metabolic fate of non-esterified fatty acids in isolated hepatocytes from newborn and young pigs. Evidence for a limited capacity for oxidation and increased capacity for esterification. Biochemical Journal 212, 9397.Google Scholar
Pégorier, JP, Ferré, P, Girard, J 1977. The effects of inhibition of fatty acid oxidation in suckling newborn rats. Biochemical Journal 166, 631634.Google Scholar
Priore, P, Giudetti, AM, Natali, F, Gnoni, GV, Geelen, MJ 2007. Metabolism and short-term metabolic effects of conjugated linoleic acids in rat hepatocytes. Biochimica et Biophysica Acta 1771, 12991307.Google Scholar
Purushotham, A, Shrode, GE, Wendel, AA, Liu, LF, Belury, MA 2007. Conjugated linoleic acid does not reduce body fat but decreases hepatic steatosis in adult Wistar rats. Journal of Nutritional Biochemistry 18, 676684.CrossRefGoogle Scholar
Soler-Argilaga, C, Russell, RL, Heimberg, M 1977. Reciprocal relationship between uptake of Ca++ and biosynthesis of glycerolipids from sn-glycerol-3-phosphate by rat liver microsomes. Biochemical and Biophysical Research Communications 78, 10531059.CrossRefGoogle ScholarPubMed
Sparks, JD, Sparks, CE 1994. Insulin regulation of triacylglycerol-rich lipoprotein synthesis and secretion. Biochimica et Biophysica Acta 1215, 932.CrossRefGoogle ScholarPubMed
Stangl, GI, Müller, H, Kirchgessner, M 1999. Conjugated linoleic acid effects on circulating hormones, metabolites and lipoproteins, and its proportion in fasting serum and erythrocyte membranes of swine. European Journal of Nutrition 38, 271277.Google Scholar
Svedberg, J, Björntorp, P, Smith, U, Lönnroth, P 1990. Free-fatty acid inhibition of insulin binding, degradation, and action in isolated rat hepatocytes. Diabetes 39, 570574.Google Scholar
Thiel-Cooper, RL, Parrish, JRFC, Sparks, JC, Wiegand, BR, Ewan, RC 2001. Conjugated linoleic acid changes swine performance and carcass composition. Journal of Animal Science 79, 18211828.Google Scholar
Tsuboyama-Kasaoka, N, Takahashi, M, Tanemura, K, Kim, HJ, Tange, T, Okuyama, H, Kasai, M, Ikemoto, S, Ezaki, O 2000. Conjugated linoleic acid supplementation reduces adipose tissue by apoptosis and develops lipodystrophy in mice. Diabetes 49, 15341542.CrossRefGoogle ScholarPubMed
Williamson, JR, Browning, ET, Scholz, R 1969. Control of mechanisms of gluconeogenesis and ketogenesis. Effects of oleate on gluconeogenesis in perfused rat liver. Journal of Biological Chemistry 244, 46074616.Google Scholar
Witters, LA, Trasko, CS 1979. Regulation of hepatic free fatty acid metabolism by glucagon and insulin. American Journal of Physiology 237, E23E29.Google Scholar
Yamamoto, K, Fukuda, N, Fukui, M, Kai, Y, Ikeda, H, Sakai, T 1996. Increased secretion of triglyceride and cholesterol following inhibition of long-chain fatty acid oxidation in rat liver. Annals of Nutrition and Metabolism 40, 157164.Google Scholar
Youn, JH, Youn, MS, Bergman, RN 1986. Synergism of glucose and fructose in net glycogen synthesis in perfused rat livers. Journal of Biological Chemistry 261, 1596015969.CrossRefGoogle ScholarPubMed