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Amino Acids

, Volume 37, Issue 4, pp 751–760 | Cite as

Colonic luminal ammonia and portal blood l-glutamine and l-arginine concentrations: a possible link between colon mucosa and liver ureagenesis

  • Mamy Eklou-Lawson
  • Françoise Bernard
  • Nathalie Neveux
  • Catherine Chaumontet
  • Cécile Bos
  • Anne-Marie Davila-Gay
  • Daniel Tomé
  • Luc Cynober
  • François Blachier
Original Article

Abstract

The highest ammonia concentration in the body is found in the colon lumen and although there is evidence that this metabolite can be absorbed through the colonic epithelium, there is little information on the capacity of the colonic mucosa to transfer and metabolize this compound. In the present study, we used a model of conscious pig with a canula implanted into the proximal colon to inject endoluminally increasing amounts of ammonium chloride and to measure during 5 h the kinetics of ammonia and amino acid concentration changes in the portal and arterial blood. By injecting as a single dose from 1 to 5 g ammonia into the colonic lumen, a dose-related increase in ammonia concentration in the portal blood was recorded. Ammonia concentration remained unchanged in the arterial blood except for the highest dose tested, i.e. 5 g which thus apparently exceeds the hepatic ureagenesis capacity. By calculating the apparent net ammonia absorption, it was determined that the pig colonic epithelium has the capacity to absorb 4 g ammonia. Ammonia absorption through the colonic epithelium was concomitant with increase of l-glutamine and l-arginine concentrations in the portal blood. This coincided with the expression of both glutamate dehydrogenase and glutamine synthetase in isolated colonic epithelial cells. Since l-glutamine and l-arginine are known to represent activators for liver ureagenesis, we propose that increased portal concentrations of these amino acids following increased ammonia colonic luminal concentration represent a metabolic link between colon mucosa and liver urea biosynthesis.

Keywords

Ammonia absorption Pig colon mucosa 

Notes

Acknowledgments

The authors wish to thank Jean-Claude Bernardin, Pierre Vaugelade and Christian Poirier for their expert contribution in animal experimentation. Mamy Eklou-Lawson was a recipient of a grant from the French Ministry of Foreign Affairs (Service de Coopération et d’Action Culturelle SCAC-French Embassy in Togo).

References

  1. Ardawi MS, Newsholme EA (1985) Fuel utilization in colonocytes of the rat. Biochem J 231:713–719PubMedGoogle Scholar
  2. Birkett A, Muir J, Phillips J, Jones G, O’Dea K (1996) Resistant starch lowers fecal concentrations of ammonia and phenols in humans. Am J Clin Nutr 63:766–772PubMedGoogle Scholar
  3. Blachier F, Mariotti F, Huneau JF, Tome D (2007) Effects of amino acid-derived luminal metabolites on the colonic epithelium and physiopathological consequences. Amino Acids 33:547–562PubMedCrossRefGoogle Scholar
  4. Bos C, Airinei G, Mariotti F, Benamouzig R, Berot S, Evrard J, Fenart E, Tome D, Gaudichon C (2007) The poor digestibility of rapeseed protein is balanced by its very high metabolic utilization in humans. J Nutr 137:594–600PubMedGoogle Scholar
  5. Chacko A, Cummings JH (1988) Nitrogen losses from the human small bowel: obligatory losses and the effect of physical form of food. Gut 29:809–815PubMedCrossRefGoogle Scholar
  6. Closs EI, Albritton LM, Kim JW, Cunningham JM (1993) Identification of a low affinity, high capacity transporter of cationic amino acids in mouse liver. J Biol Chem 268:7538–7544PubMedGoogle Scholar
  7. Codina J, Pressley TA, DuBose TD Jr (1999) The colonic H+, K+-ATPase functions as a Na+-dependent K+(NH4 +)-ATPase in apical membranes from rat distal colon. J Biol Chem 274:19693–19698PubMedCrossRefGoogle Scholar
  8. Cohen RM, Stephenson RL, Feldman GM (1988) Bicarbonate secretion modulates ammonium absorption in rat distal colon in vivo. Am J Physiol 254:F657–F667PubMedGoogle Scholar
  9. Cremin JD Jr, Fitch MD, Fleming SE (2003) Glucose alleviates ammonia-induced inhibition of short-chain fatty acid metabolism in rat colonic epithelial cells. Am J Physiol 285:G105–G114Google Scholar
  10. Darcy-Vrillon B, Morel MT, Cherbuy C, Bernard F, Posho L, Blachier F, Meslin JC, Duee PH (1993) Metabolic characteristics of pig colonocytes after adaptation to a high fiber diet. J Nutr 123:234–243PubMedGoogle Scholar
  11. Darcy-Vrillon B, Cherbuy C, Morel MT, Durand M, Duee PH (1996) Short chain fatty acid and glucose metabolism in isolated pig colonocytes: modulation by NH4 +. Mol Cell Biochem 156:145–151PubMedCrossRefGoogle Scholar
  12. Darragh AJ, Cranwell PD, Moughan PJ (1994) Absorption of lysine and methionine from the proximal colon of the piglet. Br J Nutr 71:739–752PubMedCrossRefGoogle Scholar
  13. De Preter V, Coopmans T, Rutgeerts P, Verbeke K (2006) Influence of long-term administration of lactulose and Saccharomyces boulardii on the colonic generation of phenolic compounds in healthy human subjects. J Am Coll Nutr 25:541–549PubMedGoogle Scholar
  14. Eisenberg D, Gill HS, Pfluegl GM, Rotstein SH (2000) Structure–function relationships of glutamine synthetases. Biochim Biophys Acta 1477:122–145PubMedGoogle Scholar
  15. Evenepoel P, Claus D, Geypens B, Hiele M, Geboes K, Rutgeerts P, Ghoos Y (1999) Amount and fate of egg protein escaping assimilation in the small intestine of humans. Am J Physiol 277:G935–G943PubMedGoogle Scholar
  16. Geypens B, Claus D, Evenepoel P, Hiele M, Maes B, Peeters M, Rutgeerts P, Ghoos Y (1997) Influence of dietary protein supplements on the formation of bacterial metabolites in the colon. Gut 41:70–76PubMedGoogle Scholar
  17. Gibson JA, Sladen GE, Dawson AM (1976) Protein absorption and ammonia production: the effects of dietary protein and removal of the colon. Br J Nutr 35:61–65PubMedCrossRefGoogle Scholar
  18. Govers MJ, Gannon NJ, Dunshea FR, Gibson PR, Muir JG (1999) Wheat bran affects the site of fermentation of resistant starch and luminal indexes related to colon cancer risk: a study in pigs. Gut 45:840–847PubMedCrossRefGoogle Scholar
  19. Handlogten ME, Hong SP, Zhang L, Vander AW, Steinbaum ML, Campbell-Thompson M, Weiner ID (2005) Expression of the ammonia transporter proteins Rh B glycoprotein and Rh C glycoprotein in the intestinal tract. Am J Physiol 288:G1036–G1047Google Scholar
  20. Haussinger D (1990) Liver glutamine metabolism. JPEN 14:56S–62SGoogle Scholar
  21. Heijnen ML, Deurenberg P, van Amelsvoort JM, Beynen AC (1997) Retrograded (RS3) but not uncooked (RS2) resistant starch lowers fecal ammonia concentrations in healthy men. Am J Clin Nutr 65:167–169PubMedGoogle Scholar
  22. Hume ID, Karasov WH, Darken BW (1993) Acetate, butyrate and proline uptake in the caecum and colon of prairie voles (Microtus ochrogaster). J Exp Biol 176:285–297PubMedGoogle Scholar
  23. Inoue H, Jackson SD, Vikulina T, Klein JD, Tomita K, Bagnasco SM (2004) Identification and characterization of a Kidd antigen/UT-B urea transporter expressed in human colon. Am J Physiol 287:C30–C35CrossRefGoogle Scholar
  24. James LA, Lunn PG, Middleton S, Elia M (1998) Distribution of glutaminase and glutamine synthetase activities in the human gastrointestinal tract. Clin Sci (Lond) 94:313–319Google Scholar
  25. Jonbloed AW, Mroz Z (1997) Intestinal absorption and secretion of minerals along the digestive tract of pigs. In: Laplace JP, Février C, Barbeau A (eds) Digestive physiology in pigs. European Association of Animal Production, Rome, Italy, pp 288–299Google Scholar
  26. Kramer P (1966) The effect of varying sodium loads on the ileal excreta of human ileostomized subjects. J Clin Inves 45:1710–1718CrossRefGoogle Scholar
  27. Le Bacquer O, Nazih H, Blottiere H, Meynial-Denis D, Laboisse C, Darmaun D (2001) Effects of glutamine deprivation on protein synthesis in a model of human enterocytes in culture. Am J Physiol 281:G1340–G1347Google Scholar
  28. Leigh JA, Dodsworth JA (2007) Nitrogen regulation in bacteria and archaea. Ann Rev Microbiol 61:349–377CrossRefGoogle Scholar
  29. Lin HC, Visek WJ (1991a) Colon mucosal cell damage by ammonia in rats. J Nutr 121:887–893PubMedGoogle Scholar
  30. Lin HC, Visek WJ (1991b) Large intestinal pH and ammonia in rats: dietary fat and protein interactions. J Nutr 121:832–843PubMedGoogle Scholar
  31. Meijer AJ, Lamers WH, Chamuleau RA (1990) Nitrogen metabolism and ornithine cycle function. Physiol Rev 70:701–748PubMedGoogle Scholar
  32. Meynial-Denis D, Mignon M, Miri A, Imbert J, Aurousseau E, Taillandier D, Attaix D, Arnal M, Grizard J (1996) Glutamine synthetase induction by glucocorticoids is preserved in skeletal muscle of aged rats. Am J Physiol 271:E1061–E1066PubMedGoogle Scholar
  33. Moran BJ, Jackson AA (1990) 15N-urea metabolism in the functioning human colon: luminal hydrolysis and mucosal permeability. Gut 31:454–457PubMedCrossRefGoogle Scholar
  34. Mortensen PB, Clausen MR (1996) Short-chain fatty acids in the human colon: relation to gastrointestinal health and disease. Scand J Gastroenterol 216:132–148CrossRefGoogle Scholar
  35. Mouille B, Morel E, Robert V, Guihot-Joubrel G, Blachier F (1999) Metabolic capacity for l-citrulline synthesis from ammonia in rat isolated colonocytes. Biochim Biophys Acta 1427:401–407PubMedGoogle Scholar
  36. Mouille B, Robert V, Blachier F (2004) Adaptative increase of ornithine production and decrease of ammonia metabolism in rat colonocytes after hyperproteic diet ingestion. Am J Physiol 287:G344–G351Google Scholar
  37. Raabe W (1990) Effects of NH4+ on the function of the CNS. Adv Exp Med Biol 272:99–120PubMedGoogle Scholar
  38. Roediger WE (1982) Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology 83:424–429PubMedGoogle Scholar
  39. Singh SK, Binder HJ, Geibel JP, Boron WF (1995) An apical permeability barrier to NH3/NH4 + in isolated, perfused colonic crypts. Proc Natl Acad Sci USA 92:11573–11577PubMedCrossRefGoogle Scholar
  40. Smiddy FG, Gregory SD, Smith IB, Goligher JC (1960) Faecal loss of fluid, electrolytes, and nitrogen in colitis before and after ileostomy. Lancet 1:14–19PubMedCrossRefGoogle Scholar
  41. Smith MW, James PS (1976) Amino acid transport by the helicoidal colon of the new-born pig. Biochim Biophys Acta 419:391–394PubMedCrossRefGoogle Scholar
  42. Smith CP, Potter EA, Fenton RA, Stewart GS (2004) Characterization of a human colonic cDNA encoding a structurally novel urea transporter, hUT-A6. Am J Physiol 287:C1087–C1093CrossRefGoogle Scholar
  43. Stewart GS, Fenton RA, Thevenod F, Smith CP (2004) Urea movement across mouse colonic plasma membranes is mediated by UT-A urea transporters. Gastroenterology 126:765–773PubMedCrossRefGoogle Scholar
  44. Summerskill WH, Wolpert E (1970) Ammonia metabolism in the gut. Am J Clin Nutr 23:633–639PubMedGoogle Scholar
  45. Vaugelade P, Posho L, Darcy-Vrillon B, Bernard F, Morel MT, Duee PH (1994) Intestinal oxygen uptake and glucose metabolism during nutrient absorption in the pig. Proc Soc Exp Biol Med 207:309–316PubMedGoogle Scholar
  46. Vince AJ, Burridge SM (1980) Ammonia production by intestinal bacteria: the effects of lactose, lactulose and glucose. J Med Microbiol 13:177–191PubMedCrossRefGoogle Scholar
  47. Watford M, Darcy-Vrillon B, Duee PH (2000) Dietary glutamine suppresses endogenous glutamine turnover in the rat. Metab Clin Exp 49:141–145PubMedGoogle Scholar
  48. Wolpert E, Phillips SF, Summerskill WH (1971) Transport of urea and ammonia production in the human colon. Lancet 2:1387–1390PubMedCrossRefGoogle Scholar
  49. Wrong O, Metcalfe-Gibson A, Morrison RB, Ng ST, Howard AV (1965) In vivo dialysis of faeces as a method of stool analysis. I. Technique and results in normal subjects. Clin Sci 28:357–375PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Mamy Eklou-Lawson
    • 1
  • Françoise Bernard
    • 2
  • Nathalie Neveux
    • 3
    • 4
  • Catherine Chaumontet
    • 1
  • Cécile Bos
    • 1
  • Anne-Marie Davila-Gay
    • 1
  • Daniel Tomé
    • 1
  • Luc Cynober
    • 3
    • 4
  • François Blachier
    • 1
  1. 1.INRA, CNRH-IdF, AgroParisTech, UMR 914 Nutrition Physiology and Ingestive BehaviorParisFrance
  2. 2.INRA, Unité d’Ecologie et de Physiologie du Système DigestifJouy-en-JosasFrance
  3. 3.APHP Laboratoire de BiochimieHopital Hôtel-DieuParisFrance
  4. 4.Laboratoire de Biologie de la Nutrition, Faculté de PharmacieUniversité Paris DescartesParisFrance

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