Abstract
Aim:
Transformation and possible metabolic effects of extracellular NAD+ were investigated in the livers of mice (Mus musculus; Swiss strain) and rats (Rattus novergicus; Holtzman and Wistar strains).
Methods:
The livers were perfused in an open system using oxygen-saturated Krebs/Henseleit-bicarbonate buffer (pH 7.4) as the perfusion fluid. The transformation of NAD+ was monitored using high-performance liquid chromatography.
Results:
In the mouse liver, the single-pass metabolism of 100 μmol/L NAD+ was almost complete; ADP-ribose and nicotinamide were the main products in the outflowing perfusate. In the livers of both Holtzman and Wistar rats, the main transformation products were ADP-ribose, uric acid and nicotinamide; significant amounts of inosine and AMP were also identified. On a weight basis, the transformation of NAD+ was more efficient in the mouse liver. In the rat liver, 100 μmol/L NAD+ transiently inhibited gluconeogenesis and oxygen uptake. Inhibition was followed by a transient stimulation. Inhibition was more pronounced in the Wistar strain and stimulation was more pronounced in the Holtzman strain. In the mouse liver, no clear effects on gluconeogenesis and oxygen uptake were found even at 500 μmol/L NAD+.
Conclusion:
It can be concluded that the functions of extracellular NAD+ are species-dependent and that observations in one species are strictly valid for that species. Interspecies extrapolations should thus be made very carefully. Actually, even variants of the same species can demonstrate considerably different responses.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Liersch M, Grotelüschen H, Decker K. NAD permeation into the liver cell. Hoppe-Seyler's Z Physiol Chem 1971; 352: 267–74.
Ziegler M. New functions of a long-known molecule. Emerging roles of NAD in cellular signaling. Eur J Biochem 2000; 267: 1550–64.
Rusinko N, Lee HC. Widespread occurrence in animal tissues of an enzyme catalyzing the conversion of NAD+ into a cyclic metabolite with intracellular Ca2+-mobilizing activity. J Biol Chem 1989; 264: 11725–31.
Chini EN, Klener P Jr, Beers KW, Chini CCS, Grande JP, Dousa TP . Cyclic ADP-ribose metabolism in rat kidney: high capacity for synthesis in glomeruli. Kidney Int 1997; 51: 1500–6.
Khoo KM, Chang CF. Localization of plasma membrane CD38 is domain specific in rat hepatocyte. Arch Biochem Biophys 2000; 373: 35–43.
Broetto-Biazon AC, Kangussu M, Sa-Nakanishi AB, Lopez CH, Constantin J, Kelmer-Bracht AM et al. Transformation products of extracellular NAD+ in the rat liver: kinetics of formation and metabolic action. Mol Cell Biochem 2008; 307: 41–50.
Haag F, Adriouch S, Braβ A, Jung C, Möller S, Scheuplein F, et al. Extracellular NAD and ATP: partners in immune cell modulation. Purinergic Signal 2007; 3: 71–81.
Broetto-Biazon AC, Bracht A, Ishii-Iwamoto EL, Silva VM, Kelmer-Bracht AM. The action of extracellular NAD+ on Ca2+ efflux, hemodynamics and some metabolic parameters in the isolated perfused rat liver. Eur J Pharmacol 2004; 484: 291–301.
Martins AG, Constantin J, Bracht F, Kelmer-Bracht AM, Bracht A. The action of extracellular NAD+ on gluconeogenesis in the perfused rat liver. Mol Cell Biochem 2006; 286: 115–24.
Cockayne DA, Muchamuel T, Grimaldi JC, Muller-Steffner H, Randall TD, Lund FE, et al. Mice deficient for the ecto-nicotinamide adenine dinucleotide glycohydrolase CD38 exhibit altered humoral immune responses. Blood 1998; 92: 1324–33.
Ceni C, Pochon N, Villaz M, Muller-Steffner H, Schuber F, Baratier J, et al. The CD38-independent ADP-ribosyl cyclase from mouse brain synaptosomes: a comparative study of neonate and adult brain. Biochem J 2006; 395: 417–26.
Scholz R, Bücher T. Hemoglobin-free perfusion of rat liver. In: Chance B, Estabrook RW, Williamson JR, editors. Control of energy metabolism. New York: Academic Press; 1965. p393–414.
Bergmeyer HU, Bernt E. Determination of glucose with glucose oxidase and peroxidase. In: Bergmeyer HU, editor. Methods of enzymatic analysis. Weinheim-London: Verlag Chemie-Academic Press; 1974. p1205–15.
Sa-Nakanishi AB, Bracht F, Yamamoto NS, Padilha F, Kelmer-Bracht AM, Bracht A. The action of extracellular NAD+ in the liver of healthy and tumor-bearing rats: Model analysis of the tumor-induced modifed response. Exp Mol Pathol 2008; 84: 218–25.
Aksoy P, White TA, Thompson M, Chini EM. Regulation of intracellular levels of NAD: a novel role for CD38. Biochem Biophys Res Commun 2006; 345: 1386–92.
Amar-Costesec A, Prado-Figueroa M, Beaufay H, Nagelkerke JF, van Berkel TJ. Analytical study of microsomes and isolated subcellular membranes from rat liver: IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells. J Cell Biol 1985; 100: 89–97.
Bersani-Amado CA, Barbuto JA, Jancar S. Comparative study of adjuvant induced arthritis in susceptible and resistant strains of rats. I. Effect of cyclophosphamide. J Rheumatol 1990; 17: 149–52.
Banik RK, Kasai M, Mizumura K. Reexamination of the difference in susceptibility to adjuvant-induced arthritis among LEW/Crj, Slc/Wistar/ST and Slc/SD rats. Exp Anim 2002; 51: 197–201.
Weinberg ML, Moreira E, Weinberg J. Arachidonic acid products-mediated contraction induced by bradykinin in relaxed mesenteric arterial rings from Holtzman rats. Eur J Pharmacol 1997; 320: 145–50.
Dwyer SD, Meyers KM. Rat platelet aggregation: strain and stock variations. Thromb Res 1986; 42: 49–53.
Lee HC. Structure and enzymatic functions of human CD38. Mol Med 2006; 12: 317–23.
Acknowledgements
This work has received financial support from the Fundação Araucária (Paraná) and the Conselho Nacional de Desenvolvimento Científico and Tecnológico (CNPq).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Broetto-Biazon, A., Bracht, F., Bracht, L. et al. Transformation and action of extracellular NAD+ in perfused rat and mouse livers. Acta Pharmacol Sin 30, 90–97 (2009). https://doi.org/10.1038/aps.2008.7
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2008.7
