Summary
1. Polymorphonuclear leucocytes from normal and diabetic subjects and from normal and alloxan diabetic rats were incubated with 14C-glucose, and allowed to phagocytize. — 2. The major 14CO2 production originated from the pentose cycle. Approx. one third of 14CO2 was formed by decarboxylation of pyruvate, whereas Krebs cycle activity was minimal. — 3. The pentose cycle metabolized 0.1–0.5% of the phosphorylated glucose, and phagocytosis increased this fraction 2–3 fold. No difference was found between normal and diabetic cells. — 4. The major endproduct of glucose metabolism was lactic acid. The randomization of 14C of glucose was not consistent with the sole operation of the oxidative or non-oxidative parts of the pentose cycle, but indicated the participation of a symmetrical C3 intermediate, i.e. dihydroxy-acetone, in the metabolism of part of the glucose carbons. — 5. Very small amounts of 14C from glucose were found in aminoacids, metabolic acids not extractable with ether, and lipids. Appreciable amounts of 14C were, however, found in a compound (A), part of which precipitated as glyceroltribenzoate, and a compound (B), which presumably was of protein nature. The incorporation of 14C in compound A from normal leucocytes decreased during phagocytosis, and was insignificant in human diabetic cells. The incorporation of 14C from [1-14C]- and [2-14C]-, but not from [6-14C]-glucose, into compound B was greatly stimulated in normal cells during phagocytosis, but not so in diabetic leucocytes.
Résumé
1. Des leucocytes polynucléaires de sujets normaux et diabétiques et de rats normaux et rendus diabétiques par l'alloxane ont été incubés avec du 14C-glucose et mis à phagocyter. — 2. La majeure production de 14CO2 venait du cycle des pentoses. Approximativement un tiers du 14CO2 était formé par la decarboxylation du pyruvate, tandis que l'activité du cycle de Krebs était minime. — 3. Le cycle des pentoses métabolisait 0.1–0.5% du glucose phosphprylé, et la phagocytose augmentait 2–3 fois cette fraction. On n'a trouvé aucune différence entre les cellules normales et les cellules de diabétiques. — 4. Le produit final principal du métabolisme du glucose était l'acide lactique. La dispersion du 14C du glucose n'était pas compatible avec le seul fonctionnement des parties oxydatives et non-oxydatives du cycle des pentoses, mais indiquait la participation d'un intermédiaire symétrique en C3, c-à-d la dihydroxyacétone, dans le métabolisme d'une partie des carbones du glucose. — 5. De très petites quantités de 14C du glucose ont été trouvées dans les acides aminés, les acides métaboliques non-extractibles par l'éther et dans les lipides. Cependant on a trouvé des quantités appréciables de 14C dans un composé (A) dont une partie précipitait comme tribenzoate de glycérol, et un composé (B) qui était probablement de nature protéique. L'incorporation de 14C dans le composé A de leucocytes normaux diminuait au cours de la phatocytose et était insignifiante dans les cellules de diabétiques humains. — L'incorporation de 14C du glucose [1-14C]- et [2-14C]-, mais non du glucose [6-14C] dans le composé B était fortement stimulée dans les cellules normales au cours de la phagocytose, mais pas autant dans les leucocytes de diabétiques.
Zusammenfassung
1. Polymorphkernige Leukozyten von Normalpersonen und Diabetikern und von normalen und alloxandiabetischen Ratten wurden mit 14C-Glucose inkubiert und zur Phagozytose angeregt. — 2. Der überwiegende Teil der 14CO2 Produktion stammte aus dem Pentosezyklus. Etwa ein Drittel des 14CO2 wurde bei der Decarboxylierung von Pyruvat gebildet, während die Aktivität des Krebs-Zyklus nur sehr gering war. — 3. Den Pentose-Phosphat-Zyklus durchliefen 0.1 bis 0.5% der phosphorylierten Glucose. Phagozytose erhöhte diesen Anteil auf das 2- bis 3-fache. Zwischen normalen und diabetischen Zellen fanden sich dabei keine Unterschiede. — 4. Milchsäure stellte das Hauptendprodukt des Glucose-stoffwechsels dar. Die Verteilung des 14C aus der Glucose spiegelte nicht nur die Einwirkungen des oxydativen und des nicht-oxydativen Anteils des Pentose-Phosphat-Zyklus wider, sondern deutete auch auf die Beteiligung eines symmetrischen C3 Zwischenproduktes, nämlich Dihydroxyaceton, beim Stoffwechsel eines Teils des Glucose-Kohlenstoffs hin. — 5. Nur sehr kleine Mengen des 14C aus Glucose fanden sich in Aminosäuren, nicht mit Äther extrahierbaren Stoffwechsel-Säuren und Lipiden. Hingegen ließen sich beträchtliche Anteile des 14C in einem Stoffgemisch (A) nachweisen, von dem ein Teil als Glycerin-Tribenzoat präzipitierte, und in einem Gemisch (B), das wahrscheinlich Eiweißcharakter besitzt. Die Einlagerung von 14C in das Gemisch A durch normale Leukozyten nahm während der Phagozytose ab und war unbedeutend bei diabetischen Zellen. Der Einbau von 14C aus [1-14C]- und [2-14C]-, nicht aber aus [6-14C]-Glucose in das Gemisch B war bei normalen Zellen während der Phagozytose erheblich gesteigert, jedoch nicht bei diabetischen Zellen.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Beck, W.S.: Occurrence and control of the phospho-gluconate oxidation pathway in normal and leukemic leucocytes. J. biol. Chem. 232, 271–283 (1958).
Berger, R.R., and M.L. Karnowsky: Biochemical basis of phagocytosis. V. Effect of phagocytosis on cellular uptake of extracellular fluid, and on the intracellular pool of L-α-glycerophosphate. Fed. Proc. 25, 840–845 (1966).
Busch, H., R.B. Hurlbert, and V.R. Potter: Anion exchange chromatography of acids of the citric acid cycle. J. biol. Chem. 196, 717–727 (1952).
Elsbach, P.: Composition and synthesis of lipids in resting and phagocytizing leukocytes. J. exp. Med. 110, 969–980 (1959).
Esmann, V.: Carbohydrate metabolism and respiration in leukocytes from normal and diabetic subjects, p. 40, 71, 89, 108, 118 Aarhus: Universitetsforlaget 1962.
—: Effect of cell concentration on the metabolism of normal and diabetic leukocytes in vitro. Metabolism 13, 354–360 (1964).
—: Effect of insulin on human leukocytes. Diabetes 12, 545–549 (1963).
- The metabolism of glycerol and glyceraldehyde in rat polymorphonuclear leukocytes. Acta chem. scand., 1968, in press.
—, C.J. Hedeskov, and M. Rosell Perez: UDPGlucose-glucan glucosyl-transferase activity of polymorphonuclear leucocytes from diabetic subjects. Diabetologia 4, 181–187 (1968).
—, E.P. Noble, and R. Stjernholm: Carbohydrate metabolism in leukocytes. VI. The metabolism of mannose and fructose in polymorphonuclear leukocytes of rabbit. Acta chem. scand. 19, 1672–76 (1965).
Evans, W.H., and M.L. Karnowsky: The biochemical basis of phagocytosis. IV. Some aspects of carbohydrate metabolism during phagocytosis. Biochemistry 1, 159–166 (1962).
Hedeskov, C.J., and V. Esmann: Major metabolic pathways of glucose in normal human lymphocytes and the effect of cortisol. Bioch. biophys. Acta 148, 372–383 (1968).
Hennes, A.R., and K. Awai: Studies of incorporation of radioactivity into lipids by human blood. III. Abnormal incorporation of acetate-14C into fatty acids by whole blood and platelets from nonketotic and insulin-dependent diabetics. Metabolism 14, 487–499 (1965).
—: IV. Abnormal incorporation of acetate- [1-14C] into fatty acids by whole blood and platelets from insulin independent diabetics. Diabetes 14, 708–715 (1965).
Katz, J., and R. Rognstad: The labelling of pentose-P from glucose-14C and estimation of the rates of transaldolase, transketolase, the contribution of the pentose cycle and ribose phosphate synthesis. Biochemistry 6, 2227–2247 (1967).
—, and H.G. Wood: The use of 14CO2 yields from glucose-1- and -[6-14C] for the evaluation of the pathways of glucose metabolism. J. biol. Chem. 238, 517–523 (1963).
Morton, D.J., J.F. Moran, N. Dimitrov, and W. Falor: Metabolic stimulation and phagocytosis by leukocytes. Fed. Proc. 25, 762 (1966), Abstr. 3240.
Noble, E.P., R. Stjernholm, and L. Ljungdahl: Carbohydrate metabolism in leukocytes. III. Carbon dioxide incorporation in the rabbit polymorphonuclear leukocyte. Biochim. biophys. Acta. 49, 593–595 (1961).
— —, and A.S. Weisberger: Carbohydrate metabolism in the leukocytes. I. The pathway of two- and three-carbon compounds in the rabbit polymorphonuclear leukocyte. J. biol. Chem. 235, 1261–64 (1960).
Pitkänen, E., and E.A. Nikkilä: Enzyme pattern of human leukocytes in diabetes and thyroid diseases. Ann. Med. intern. Fenn. 49, 275–281 (1960).
Sbarra, A.J., and M.L. Karnowsky: The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J. biol. Chem. 234, 1355–62 (1959).
Selvaraj, R.J., and A.J. Sbarra: Phagocytosis inhibition and reversal II. Possible role of pyruvate as an alternative source of energy for particle uptake by guinea-pig leukocytes. Biochim. biophys. Acta 127, 159–171 (1966).
Stjernholm, R., and E.P. Noble: Carbohydrate metabolism in leukocytes. II. The pathway of ribose and xylose metabolism in the rabbit polymorphonuclear leukocyte. J. biol. Chem. 236, 614–616 (1961).
— —: IV. The metabolism of glucose and galactose in polymorphonuclear leukocytes from rabbit. J. biol. Chem. 236, 3093–3096 (1961).
— —: V. The metabolism of five-carbon substrates in polymorphonuclear leukocytes of rabbit. Arch. biochem. 100, 200–204 (1963).
—, and H.G. Wood: Glycerol dissimilation and occurrence of a symmetrical three-carbon intermediate in propionic acid formentation. J. biol. Chem. 235, 2757–2761 (1960).
Swim, E.H., and L.O. Krampitz: Acetic acid oxidation by Escherichia Coli: Evidence for the occurrence of a tricarboxylic acid cycle. J. Bact. 67, 419–425 (1954).
Wood, H.G., and J. Katz: The distribution of 14C in the hexose phosphates and the effect of recycling in the pentose cycle. J. biol. Chem. 233, 1279–1282 (1958).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Esmann, V. The metabolism of [1-14C]-, [2-14C]-, [3,4-14C]-, and [6-14C]- glucose in normal and diabetic polymorphonuclear leukocytes and during phagocytosis. Diabetologia 4, 188–194 (1968). https://doi.org/10.1007/BF00430094
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00430094