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Metabolic differences in hepatocytes of obese and lean pigs

Published online by Cambridge University Press:  15 July 2014

L. González-Valero ,
J. M. Rodríguez-López ,
M. Lachica  and
I. Fernández-Fígares
L. González-Valero
Affiliation:
Department of Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Camino del Jueves s/n, 18100 Armilla, Granada, Spain
J. M. Rodríguez-López
Affiliation:
Department of Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Camino del Jueves s/n, 18100 Armilla, Granada, Spain
M. Lachica
Affiliation:
Department of Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Camino del Jueves s/n, 18100 Armilla, Granada, Spain
I. Fernández-Fígares*
Affiliation:
Department of Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Camino del Jueves s/n, 18100 Armilla, Granada, Spain
*
† E-mail: [email protected]
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Abstract

There are important differences in terms of metabolic activity, energy utilization and capacity of protein and fat deposition when Iberian and modern pigs are compared. Primary culture of hepatocytes was used to evaluate hepatic function and sensitivity to hormones between breeds without the interference of circulating blood factors. Hepatocytes were isolated from pure Iberian (n=10) and Landrace (n=8) pigs of similar BW (24.5±12.1 and 32.9±6.1 kg BW, respectively), by collagenase perfusion. Monolayers were established in medium containing fetal bovine serum for 1 day and switched to serum-free medium for the remainder of the culture period. Hepatocytes were maintained in William’s E supplemented with β-mercaptoethanol (0.1 mM), glutamine (2 mM), antibiotics (gentamicin, penicillin, streptomycin and amphotericin B), dimethyl sulfoxide (1 µg/ml), dexamethasone (10−8 M), insulin (0.173 and 17.3 nM) and glucagon (0.287, 2.87 and 28.7 nM) for 24 to 48 h. Gluconeogenesis (GNG), glycogen degradation, triglycerides (TG) content and esterification, β-hydroxybutyrate (BHB) synthesis, IGF-1 synthesis, albumin and urea synthesis were determined. Iberian pigs had greater capacity of GNG than Landrace (24%, P<0.05), although no difference in glycogen degradation was found (P>0.10). TG content and esterification tended to be lower in hepatocytes from Iberian compared with Landrace pigs (12% and 31%, respectively; 0.10<P<0.05). Furthermore, addition of free fatty acids (CLA or linoleic acid, 0.2 mM) increased TG content (64%, P<0.001) although no difference between fatty acids was found. When free fatty acids were compared, a trend toward increased esterification (41%, P=0.078) was found for CLA. Although glucagon stimulated and insulin inhibited BHB synthesis, no difference between breeds was found (P>0.10). IGF-1 synthesis was diminished in hepatocytes from Iberian compared with Landrace pigs (16%, P<0.05). On the contrary, rate of albumin synthesis was greater in Iberian compared with Landrace pigs (58%, P<0.05). Finally, the capacity of urea synthesis was lower in hepatocytes of Iberian compared with Landrace pigs (37%, P<0.05). When ammonia was added to the media, urea concentration increased (648%, 1108% and 2791% when 0 mM was compared with 2.5, 5 and 10 mM, respectively). Urea synthesis increased on increasing ammonia content (55% and 325% when 0 mM was compared with 5 and 10 mM, respectively; P<0.0001). In conclusion, the genetic background accounts for important differences in protein and energy metabolism pathways found in primary culture of hepatocytes from lean and obese pigs.

Keywords

hepatocytesmetabolismpig
Type
Research Article
Information
animal , Volume 8 , Issue 11 , November 2014 , pp. 1873 - 1880
Copyright
© The Animal Consortium 2014 

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References

Ajuwon, KM, Kuske, JL, Anderson, DB, Hancock, DL, Houseknecht, KL, Adeola, O and Spurlock, ME 2003. Chronic leptin administration increases serum NEFA in the pig and differentially regulates PPAR expression in adipose tissue. The Journal of Nutritional Biochemistry 14, 576–583.CrossRefGoogle ScholarPubMed
Ballmer, PE, McNurlan, MA, Essen, P, Anderson, SE and Garlick, PJ 1995. Albumin synthesis rates measured with [2H5ring]Phenylalanine are not responsive to short-term intravenous nutrients in healthy humans. The Journal of Nutrition 125, 512–519.Google Scholar
Brameld, JM 1997. Molecular mechanisms involved in the nutritional and hormonal regulation of growth in pigs. Proceedings of the Nutrition Society 56, 607–619.Google Scholar
Buonomo, FC, Lauterio, TL, Baile, CA and Campion, DR 1987. Determination of insulin-like growth factor 1(IGF1) and IGF binding protein levels in swine. Domestic Animal Endocrinology 4, 23–31.CrossRefGoogle ScholarPubMed
Butler, AA and Le Roith, D 2001. Control of growth by the somatropic axis: growth hormone and the insulin-like growth factors have related and independent roles 1. Annual Review of Physiology 63, 141–164.Google Scholar
Carey, GB, Cheung, CW, Cohen, NS, Brusilow, S and Raijman, L 1993. Regulation of urea and citrulline synthesis under physiological conditions. The Biochemical Journal 292, 241–247.Google Scholar
Coma, J, Zimmerman, DR and Carrion, D 1995. Relationship of rate of lean tissue growth and other factors to concentration of urea in plasma of pigs. Journal of Animal Science 73, 3649–3656.Google Scholar
Conde-Aguilera, JA, Lachica, M, Nieto, R and Fernández-Fígares, I 2012. Metabolic regulation of fatty acid esterification and effects of CLA on glucose homeostasis in pig hepatocytes. Animal 6, 254–261.Google Scholar
Connell, A, Calder, AG, Anderson, SE and Lobley, GE 1997. Hepatic protein synthesis in the sheep: effect of intake as monitored by use of stable-isotope-labelled glycine, leucine and phenylalanine. The British Journal of Nutrition 77, 255–271.Google Scholar
Davis, TA, Fiorotto, ML, Beckett, PR, Burrin, DG, Reeds, PJ, Wray-Cahen, D and Nguyen, HV 2001. Differential effects of insulin on peripheral and visceral tissue protein synthesis in neonatal pigs. American Journal of Physiology Endocrinology and Metabolism 280, E770–E779.Google Scholar
Duée, PH, Pégorier, JP, Quant, PA, Herbin, C, Kohl, C and Girard, J 1994. Hepatic ketogenesis in newborn pigs is limited by low mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase activity. The Biochemical Journal 298, 207–212.CrossRefGoogle ScholarPubMed
Fabian, J, Chiba, LI, Kuhlers, DL, Frobish, LT, Nadarajah, K and McElhenney, WH 2003. Growth performance, dry matter and nitrogen digestibilities, serum profile, and carcass and meat quality of pigs with distinct genotypes. Journal of Animal Science 81, 1142–1149.Google Scholar
Fenton, JP, Roehrig, KL, Mahan, DC and Corley, JR 1985. Effect of swine weaning age on body fat and lipogenic activity in liver and adipose tissue. Journal of Animal Science 60, 190–199.CrossRefGoogle ScholarPubMed
Fernández-Fígares, I, Shannon, AE, Wray-Cahen, D and 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, 125–140.Google Scholar
Fernández-Fígares, I, Lachica, M, Nieto, R, Rivera-Ferre, MG and Aguilera, J 2007. Serum profile of metabolites and hormones in obese (Iberian) and lean (Landrace) growing gilts fed balanced or lysine deficient diets. Livestock Science 110, 73–81.Google Scholar
Fernández-Fígares, I, Lachica, M, Martín, A, Nieto, R, González-Valero, L, Rodríguez-López, JM and Aguilera, JF 2012. Impact of dietary betaine and CLA on insulin sensitivity, protein and fat metabolism of obese pigs. Animal 6, 1058–1067.Google Scholar
Go, G, Wu, G, Silvey, DT, Choi, S, Li, X and Smith, SB 2012. Lipid metabolism in pigs fed supplemental conjugated linoleic acid and/or dietary arginine. Amino Acids 43, 1713–1726.Google Scholar
Gondret, F, Ferré, P and 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, 106–113.Google Scholar
Huizenga, JR, Glips, CH and Tangerman, A 1996. The contribution of various organs to ammonia formation: a review of factors determining the arterial concentration. Annals of Clinical Biochemistry 33, 23–30.Google Scholar
Kasser, TR, Martin, RJ, Gahagan, JH and Wangsness, PJ 1981. Fasting plasma hormones and metabolites in feral and domestic newborn pigs. Journal of Animal Science 53, 420–426.Google Scholar
Katz, J, Kuwajima, M, Foster, DW and McGarry, JD 1986. The glucose paradox: new perspectives on hepatic carbohydrate metabolism. Trends in Biochemical Sciences 11, 136–140.Google Scholar
Louveau, I, Bonneau, M and Salter, DN 1991. Age-related changes in plasma porcine growth hormone (GH) profiles and insulin-like growth factor-I (IGF-I) concentrations in Large White and Meishan pigs. Reproduction Nutrition Development 31, 205–216.Google Scholar
McCusker, RH, Wangsness, PJ, Griel, LC Jr and Kavanaugh, JF 1985. Effects of feeding, fasting and refeeding on growth hormone and insulin in obese pigs. Physiology and Behavior 35, 383–388.Google Scholar
Müller, MJ, Paschen, U and Seitz, HJ 1982. Starvation-induced ketone body production in the conscious unrestrained miniature pig. Journal of Nutrition 112, 1379–1386.Google Scholar
Nieto, R, Lara, L, Barea, R, García-Valverde, R, Aguinaga, MA, Conde-Aguilera, JA and Aguilera, JF 2012. Response analysis of the Iberian pig growing from birth to 150 kg body weight to changes in protein and energy supply. Journal of Animal Science 90, 3809–3820.CrossRefGoogle ScholarPubMed
Peavy, DE, Taylor, JM and Jefferson, LS 1978. Correlation of albumin production rates and albumin mRNA levels in livers of normal, diabetic, and insulin-treated diabetic rats. Proceedings of the National Academy of Sciences of the United States of America 75, 5879–5883.CrossRefGoogle ScholarPubMed
Pégorier, JP, Duée, PH, Girard, J and Peret, J 1982. Development of gluconeogenesis in isolated hepatocytes from fasting or suckling newborn pigs. The Journal of Nutrition 112, 1038–1046.CrossRefGoogle ScholarPubMed
Pégorier, JP, Duée, PH, Girard, J and 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. The Biochemical Journal 212, 93–97.Google Scholar
Pond, WG, Yen, JT, Lindvall, RN and Hill, D 1980. Dietary alfalfa meal for genetically-obese and lean growing-pigs: effect on body-weight gain and on carcass and gastrointestinal-tract measurements and blood metabolites. Journal of Animal Science 51, 367–373.Google Scholar
Priore, P, Giudetti, AM, Natali, F, Gnoni, GV and Geelen, MJH 2007. Metabolism and short-term metabolic effects of conjugated linoleic acids in rat hepatocytes. Biochimica et Biophysica Acta 1771 (10), 1299–1307.Google Scholar
Ramsay, TG, Richards, MP, Li, CJ and Caperna, TJ 2010. IGF-I mediated inhibition of leptin receptor expression in porcine hepatocytes. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 155, 43–48.Google Scholar
Rivera-Ferre, MG, Aguilera, JF and Nieto, R 2005. Muscle fractional protein synthesis is greater in Iberian than in Landrace growing pigs fed adequate or lysine-deficient diets. The Journal of Nutrition 135, 469–478.Google Scholar
Rodríguez-López, JM, González-Valero, L, Lachica, M and Fernández-Fígares, I 2013. Growth hormone releasing factor and secretion of growth hormone in Iberian and Landrace gilts. In Energy and protein metabolism and nutrition in sustainable animal production (ed. JW Oltjen, E Kebreab and H Lapierre), pp. 293–294. Wageningen Academic Publishers, Wageningen, The Netherlands.CrossRefGoogle Scholar
Stangl, GI, Müller, H and 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, 271–277.Google Scholar
Tomas, FM, Knowles, SE, Owens, PC, Chandler, CS, Francis, GL, Read, LC and Ballard, FJ 1992. Insulin-like growth factor-I (IGF-I) and especially IGF-I variants are anabolic in dexamethasone-treated rats. The Biochemical Journal 282, 91–97.CrossRefGoogle ScholarPubMed
Wang, J, Zhu, X, Chen, C, Li, X, Gao, Y, Li, P, Zhang, Y, Long, M, Wang, Z and Liu, G 2012. Effect of insulin-like growth factor-1 (IGF-1) on the gluconeogenesis in calf hepatocytes cultured in vitro. Molecular and Cellular Biochemistry 362, 87–91.Google Scholar
Weiler, U, Claus, R, Schnoebelen-Combes, S and Louveau, I 1998. Influence of age and genotype on endocrine parameters and growth performance: a comparative study in wild boars, Meishan and Large White boars. Livestock Production Science 54, 21–31.Google Scholar
Wheelhouse, NM, Stubbs, AK, Lomax, MA, MacRae, JC and Hazlerigg, DG 1999. Growth hormone and amino acid supply interact synergistically to control insulin-like growth factor-I production and gene expression in cultured ovine hepatocytes. The Journal of Endocrinology 163, 353–361.Google Scholar
Yang, H, Pettigrew, JE, Johnston, LJ, Shurson, GC, Wheaton, JE, White, ME, Koketsu, Y, Sower, AF and Rathmacher, JA 2000. Effects of dietary lysine intake during lactation on blood metabolites, hormones, and reproductive performance in primiparous sows. Journal of Animal Science 78, 1001–1009.Google Scholar