Molecular and Cellular Biochemistry

, Volume 156, Issue 2, pp 145–151 | Cite as

Short chain fatty acid and glucose metabolism in isolated pig colonocytes: modulation by NH4+

  • Béatrice Darcy-Vrillon
  • Claire Cherbuy
  • Marie-Thérèse Morel
  • Michelle Durand
  • Pierre-Henri Duée


Short chain fatty acids (SCFA) from bacterial origin, as well as glucose from vascular origin, are among fuel substrates available to the colonic mucosa. The present work investigated the possible modulation by another bacterial metabolite, i.e. ammonia, of the capacities of colonic epithelial cells to metabolize these substrates. Viable colonocytes were isolated from the proximal colon of 40–50 kg pigs fed a standard diet and were incubated (30 min, 37°C) in the presence of a concentration range of 14C-labeled n-butyrate or acetate, or 14C-labeled glucose (5 mm), with or without NH4Cl (10 mM) addition. 14CO2 and metabolites generated were measured. Butyrate utilization resulted in a high generation of ketone bodies (acetoacetate and β-OH-butyrate), in addition to 14CO2 production. However, the net ketone body generation was significantly decreased for butyrate concentrations higher than 10 mM. In contrast to n-butyrate, acetate when given as the sole substrate got preferentially metabolized in the oxidation pathway. Acetate metabolism was not affected by NH4Cl, thus indicating that the tricarboxylic acid cycle was unchanged. Conversely, 4C02 and ketone body production from butyrate were decreased by 30% in the presence of NH4Cl, suggesting that butyrate activation or β-oxidation was diminished. Glucose utilization rate was increased by 20%, due to an increased glycolytic capacity in the presence of NH4Cl. A dose-dependent stimulation of phosphofructokinase activity by NH4+ could account for this effect. It is concluded that ammonia, whose physiological concentration is high in the colonic lumen, can modulate the metabolism of two major substrates, n-butyrate and glucose, in colonic epithelial cells.

Key words

pig colonocytes metabolism short-chain fatty acids glucose NH4+ 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Scheppach W: Effects of short chain fatty acids on gut morphology and function. Gut suppl 1: S35-S38, 1994Google Scholar
  2. 2.
    Duée PH, Darcy-Vrillon B, Blachier F, Morel MT: Fuel selection in intestinal cells. Proc Nutr Soc 54: 83–94, 1995Google Scholar
  3. 3.
    Roediger WEW: Role of anaerobic bacteria in the metabolic welfare of the colonic mucosa in man. Gut 21: 793–798, 1980Google Scholar
  4. 4.
    Roediger WEW: Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology 83: 424–129, 1982Google Scholar
  5. 5.
    Ardawi MSM, Newsholme EA: Fuel utilization in colonocytes of the rat. Biochem J 231: 713–713, 1985Google Scholar
  6. 6.
    Darcy-Vrillon B, Morel MT, Cherbuy C, Bernard F, Posho L, Blachier F, Meslin JC, Duée PH: Metabolic characteristics of pig colonocytes after adaptation to a high fiber diet. J Nutr 123: 234–243, 1993Google Scholar
  7. 7.
    Fleming SE, Fitch MD, DeVries S, Liu ML, Kight C: Nutrient utilization by cells isolated from rat jejunum. cecum and colon. J Nutr 121: 869–878, 1991Google Scholar
  8. 8.
    Kight CE, Fleming SE: Nutrient oxidation by rat intestinal epithelial cells is concentration dependent. J Nutr 123: 876–882, 1993Google Scholar
  9. 9.
    Clausen MR, Mortensen PB: Kinetic studies on the metabolism of short chain fatty acids and glucose by isolated rat colonocytes. Gastroenterology 106: 423–432, 1994Google Scholar
  10. 10.
    Roediger WEW, Duncan A, Kapaniris O, Millard S: Sulphide impairment of substrate oxidation in rat colonocytes: a biochemical basis for ulcerative colitis? Clin Sci 85: 623–627, 1993Google Scholar
  11. 11.
    Roediger WEW, Duncan A, Kapaniris O, Millard S: Reducing sulfur compounds of the colon impair colonocyte nutrition: implications for ulcerative colitis. Gastroenterology 104: 802–809, 1993Google Scholar
  12. 12.
    Roediger WEW, Kapaniris O, Millard S: Lipogenesis from n-butyrate in colonocytes. Action of reducing agent and 5-aminosalicylic acid with relevance to ulcerative colitis. Mol Cell Biochem 118: 113–118, 1992Google Scholar
  13. 13.
    MacFarlane GT, Cummings JH: The colonic flora, fermentation and large bowel digestive function. In: S.F. Phillips, J.H. Pemberton, R.G. Shorter (eds). The Large Intestine, Physiology, Pathophysiology and Disease. New York: Raven Press 1991, pp 51–92Google Scholar
  14. 14.
    Lin HC, Visek WJ: Large intestinal pH and ammonia in rats: dietary fat and protein interactions. J Nutr 121: 832–843, 1991Google Scholar
  15. 15.
    Gargallo J, Zimmerman DR: Effects of dietary cellulose and neomycin on function of the cecum of pigs. J Anim Sci 51: 121–126, 1980Google Scholar
  16. 16.
    Prior RL, Topping DC, Visek WJ: Metabolism of isolated chick small intestinal cells. Effects of ammonia and various salts. Biochemistry 13: 178–183, 1974Google Scholar
  17. 17.
    Topping DC, Visek WJ: Synthesis of macromolecules by intestinal cells incubated with ammonia. Am J Physiol 233: E341-E347, 1977Google Scholar
  18. 18.
    Roediger WEW, Truelove SC: Method of preparing isolated colonic epithelial cells (colonocytes) for metabolic studies. Gut 20: 484–488, 1979Google Scholar
  19. 19.
    Lowry OH, Rosebrough NJ, Lewis Farr A, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951Google Scholar
  20. 20.
    Bergmeyer HU: Methods of enzymatic analysis. volume 1–4 3rd ed. New York: Academic Press, 1974Google Scholar
  21. 21.
    Posho L, Darcy-Vrillon B, Morel MT, Cherbuy C, Blachier F, Duee PH: Control of glucose metabolism in newborn pig enterocytes: evidence for the role of hexokinase. Biochim Biophys Acta 1224: 213–220, 1994Google Scholar
  22. 22.
    Ardawi MSM, Newsholme EA: Maximum activities of some enzymes of glycolysis, the tricarboxylic acid cycle and ketone-body and glutamine utilization pathways in lymphocytes of the rat. Biochem J 208: 743–748, 1982Google Scholar
  23. 23.
    Sugden PH, Newsholme EA: Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenases, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates. Biochem J 150: 105–111, 1975Google Scholar
  24. 24.
    Williamson DH, Bates MW, Page MA, Krebs HA: Activities of enzymes involved in acetoacetate utilization in adult mammalian tissues. Biochem J 121: 41–47, 1971Google Scholar
  25. 25.
    Board M, Humm S, Newsholme EA: Maximum activities of key enzymes of glycolysis, glutaminolysis, pentose phosphate pathway and tricarboxylic acid cycle in normal, neoplastic and suppressed cells. Biochem J 265: 503–509, 1990Google Scholar
  26. 26.
    Tejwani GA, Ramaiah A: Properties of phosphofructokinase from the mucosa of rat jejunum and their relation to the lack of Pasteur effect. Biochem J 125: 507–514, 1971Google Scholar
  27. 27.
    Larrabee MG: The pentose cycle (Hexose monophosphate shunt). J Biol Chem 264: 15875–15879, 1989Google Scholar
  28. 28.
    Fleming SE, Arce D: Using the pig to study digestion and fermentation in the gut. In: M. Tumbleson (ed.). Swine in Biomedical Research. New York: Plenum Press 123–134, 1986Google Scholar
  29. 29.
    Clausen MR, Mortensen PB, Holtug K, Nordgaard I, Hove H, Jeppesen PB: Kinetic studies on colonocyte metabolism of short chain fatty acids and glucose in patients with ulcerative colitis. Gastroenterology, 106, A666, 1994Google Scholar
  30. 30.
    Weinman EO, Strisower EH, Chaikoff IL: Conversion of fatty acids to carbohydrate: application of isotopes to this problem and role of the Krebs cycle as a synthetic pathway. Physiol Rev 37: 252–272, 1957Google Scholar
  31. 31.
    Emmanuel B: Oxidation of butyrate to ketone bodies and CO2 in the rumen epithelium, liver, kidney, heart and lung of camel (Camelus dromedarius, sheep (Ovis aries) and goat (Carpa hircus). Comp Comp Biochem Physiol 65: 699–704Google Scholar
  32. 32.
    Cherbuy C, Darcy-Vrillon B, Morel MT, Pégorier JP, Duée PH: Effect of germ free state on the capacities of isolated rat colonocytes to metabolize n-butyrate, glucose and glutamine. Gastroenterology 109: 1890–1899, 1995Google Scholar
  33. 33.
    Endemann G, Goetz PG, Edmond J, Brunen Graber H: Lipogenesis from ketone bodies in the isolated perfused rat liver; evidence for the cytosolic activation of acetoacetate. J Biol Chem 257: 3434–3440, 1982Google Scholar
  34. 34.
    Williamson DH, Lund P, Krebs HA: The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver. Biochem J 103: 514–527, 1967Google Scholar
  35. 35.
    Khoja SM, Ardawi MSM, Abulgasim AO: Effects of starvation and streptozotocin induced diabetes on the activity of phosphofructokinase in the epithelial cells of rat colon. Biochimie 70: 721–725, 1988Google Scholar
  36. 36.
    Pritchard PJ, Lee DJW: The regulation of chick (Gallus domesticus) mucosal phosphofructokinase by ammonium and citrate ions. Int J Biochem 5: 655–660, 1974Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Béatrice Darcy-Vrillon
    • 1
  • Claire Cherbuy
    • 1
  • Marie-Thérèse Morel
    • 1
  • Michelle Durand
    • 2
  • Pierre-Henri Duée
    • 1
  1. 1.Unité d'Ecologie et de Physiologie du Système DigestifInstitut National de la Recherche AgronomiqueJouy-en-Josas, CedexFrance
  2. 2.Unité de Nutrition et Sécurité AlimentaireInstitut National de la Recherche AgronomiqueJouy-en-Josas, CedexFrance

Personalised recommendations