, Volume 161, Issue 1, pp 349–359 | Cite as

The perfused rat lung as a model for studies on the formation of surfactant and the effect of Ambroxol on this process

  • M. Post
  • J. J. Batenburg
  • E. A. J. M. Schuurmans
  • V. Oldenborg
  • A. J. van der Molen
  • L. M. G. van Golde
Original Articles


The isolated perfused rat lung was used as a model to investigate the synthesis of surfactant phospholipids from various radioactive precursors and the effect of Ambroxol, a bronchial secretolyticum, on this process. Lungs were ventilated and perfused for periods up to 5 h without detectable development of pulmonary edema. The lungs remained metabolically stable during the entire period of perfusion. Both in whole lung tissue and in the surfactant fraction the radioactive substrates incorporated predominantly into phosphatidylcholine and phosphatidylglycerol. The degree of saturation of labelled phosphatidylcholines synthesized during perfusion with [Me-14C]choline, D [U-14C]glucose, [1(3)-3H]glycerol and [1-14C]palmitate was higher in surfactant than in whole lung tissue. A delayed incorporation into surfactant phospholipids was observed for all precursors. Under the conditions employed, glucose carbon was recovered mainly in the glycerol backbone of phosphatidylcholine and phosphatidylglycerol. Compared to glucose, glycerol appeared to be a minor substrate for lung lipid formation. If the lungs were perfused after pretreatment of the rats with Ambroxol on three consecutive days, the incorporation of labelled choline and glycerol into pulmonary phospholipids was found to be enhanced. This stimulation was more pronounced in the surfactant fraction than in whole lung tissue. The stimulatory effect on the formation of surfactant lipids was smaller after pretreatment of the animals with Ambroxol for one day. The results of the present study suggest that Ambroxol may specifically stimulate the synthesis of phospholipids in the alveolar type II cells and that the drug may not only affect the formation but also the secretion of surfactant lipids by these cells.

Key words

Perfused rat lung Phosphatidylcholine Phosphatidylglycerol Surfactant Ambroxol 


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  1. 1.
    Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466–468PubMedGoogle Scholar
  2. 2.
    Bassett DJP, Hamosh M, Hamosh P, Rabinowitz J (1981) Pathways of palmitate metabolism in the isolated rat lung. Exp Lung Res 2:37–47PubMedGoogle Scholar
  3. 3.
    Batenburg JJ, Longmore WJ, Van Golde LMG (1978) The synthesis of phosphatidylcholine by adult rat lung alveolar type II epithelial cells in primary culture. Biochim Biophys Acta 529:160–170PubMedGoogle Scholar
  4. 4.
    Batenburg JJ, Post M, Oldenborg V, Van Golde LMG (1980) The perfused isolated lung as a possible model for the study of lipid synthesis by type II cells in their natural environment. Exp Lung Res 1:57–65PubMedGoogle Scholar
  5. 5.
    Batenburg JJ, Van Golde LMG (1979) Formation of pulmonary surfactant in whole lung and in isolated type II alveolar cells. In: Scarpelli EM, Cosmi EV (eds) Reviews in perinatal medicine, vol 3. Raven Press, New York, pp 73–114Google Scholar
  6. 6.
    Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917PubMedGoogle Scholar
  7. 7.
    Buechler KF, Rhoades RA (1980) Fatty acid synthesis in the perfused rat lung. Biochim Biophys Acta 619:186–195PubMedGoogle Scholar
  8. 8.
    Cerutti P, Kapanci Y (1979) Effects of metabolite VIII of bromhexine (NA 872) on type II epithelium of the lung. Respiration 37:241–251PubMedGoogle Scholar
  9. 9.
    Chang RSY, Wright K, Effros RM (1981) Role of albumin in prevention of edema in perfused rabbit lungs. J Appl Physiol 50:1065–1070PubMedGoogle Scholar
  10. 10.
    Chevalier G, Collet AJ (1972) In vivo incorporation of choline-3H, leucine-3H, galactose-3H in alveolar type II pneumocytes in relation to surfactant synthesis: a quantitative radioautographic study in mouse by electron microscopy. Anat Rec 17:289–310CrossRefGoogle Scholar
  11. 11.
    Curti PC (1972) Steigerung der Produktion der oberflächenaktiven Substanzen der Alveolen durch NA 872. Pneumologie 147:62–74CrossRefGoogle Scholar
  12. 12.
    Elphick MC, Lawlor JP (1977) Quantitative recovery of free and esterified fatty acids from thin-layer plates coated with silica. J Chromatogr 130:139–143PubMedCrossRefGoogle Scholar
  13. 13.
    Fisher AB, Dodia C, Limash J (1980) Perfusate composition and edema formation in isolated rat lungs. Exp Lung Res 1:13–21PubMedGoogle Scholar
  14. 14.
    Goerke J (1974) Lung surfactant. Biochim Biophys Acta 344:241–261PubMedGoogle Scholar
  15. 15.
    Jobe A (1977) The labeling and biological half life of phosphatidylcholine in subcellular fractions of rabbit lung. Biochim Biophys Acta 489:440–453PubMedGoogle Scholar
  16. 16.
    King RJ, Clements JA (1972) Surface active materials from dog lung. II. Composition and physiological correlations. Am J Physiol 223:715–726PubMedGoogle Scholar
  17. 17.
    Longmore WJ (1982) The isolated perfused lung as a model for studies of lung metabolism. In: Farrell PM (ed) Lung development: biological and clinical perspectives, vol 1. Academic Press, New York, London, pp 101–110Google Scholar
  18. 18.
    Lorenz U, Rüttgers H, Fux G, Kubli F (1974) Fetal pulmonary surfactant induction by bromhexine metabolite VIII. Am J Obstet Gynecol 119:1126–1128PubMedGoogle Scholar
  19. 19.
    Mason RJ, Nellenbogen J, Clements JA (1976) Isolation of disaturated phosphatidylcholine with osmium tetroxide. J Lipid Res 17:281–284PubMedGoogle Scholar
  20. 20.
    Nicholas TE, Power JHT, Barr HA (1982) The pulmonary consequences of a deep breath. Respir Physiol 49:315–324PubMedCrossRefGoogle Scholar
  21. 21.
    Oldenborg V, Van Vugt F, Van Golde LMG (1975) Composition and metabolism of phospholipids of Fasciola hepatica, the common liver fluke. Biochim Biophys Acta 398:101–110PubMedGoogle Scholar
  22. 22.
    Pande SV (1976) Liquid scintillation counting of aqueous samples using triton-containing scintillants. Anal Biochem 74:25–34PubMedCrossRefGoogle Scholar
  23. 23.
    Poorthuis BJHM, Yazaki PJ, Hostetler KY (1976) An improved two-dimensional thinlayer chromatography system for the separation of phosphatidylglycerol and its derivatives. J Lipid Res 17:433–437PubMedGoogle Scholar
  24. 24.
    Post M, Batenburg JJ, Van Golde LMG (1980) Effects of cortisol and thyroxine on phosphatidylcholine and phosphatidylglycerol synthesis by adult rat lung alveolar type II cells in primary culture. Biochim Biophys Acta 618:308–317PubMedGoogle Scholar
  25. 25.
    Postlethwait EM, Young SL (1980) Alteration of rat lung adenine nucleotide content after pulmonary edema. Lung 158:157–164PubMedCrossRefGoogle Scholar
  26. 26.
    Prevost MC, Soula G, Douste-Blazy L (1979) Biochemical modifications of pulmonary surfactant after bromhexine derivate injections. Respiration 37:215–219PubMedGoogle Scholar
  27. 27.
    Rhoades RA (1974) Net uptake of glucose, glycerol and fatty acids by the isolated perfused rat lung. Am J Physiol 226:144–149PubMedGoogle Scholar
  28. 28.
    Strehler BL (1974) Adenosine-5′-triphosphate and creatine phosphate: determination with luciferase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York London, pp 2112–2126Google Scholar
  29. 29.
    Thet LA, Clerck L, Massaro GD, Massaro D (1979) Changes in sedimentation of surfactant in ventilated excised rat lung. J Clin Invest 64:600–608PubMedCrossRefGoogle Scholar
  30. 30.
    Van Golde LMG, Van Deenen LLM (1966) The effect of dietary fat on the molecular species of lecithin from rat liver. Biochim Biophys Acta 125:496–509PubMedGoogle Scholar
  31. 31.
    Van Petten GR, Mears GJ, Taylor PJ (1978) The effect of NA 872 on pulmonary maturation in the fetal lamb and rabbit. Am J Obstet Gynecol 130:35–40PubMedGoogle Scholar
  32. 32.
    Velasquez BJ, Sépulveda J (1982) Influence of transplacental NA-872 and dexamethasone therapy on fetal and newborn mouse lung. Respiration 43:389–400PubMedCrossRefGoogle Scholar
  33. 33.
    Watkins CA, Rannels DE (1979) In situ perfusion of rat lungs: stability and effects of oxygen tension. J Appl Physiol 47:325–329PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH & Co KG 1983

Authors and Affiliations

  • M. Post
    • 1
  • J. J. Batenburg
    • 1
  • E. A. J. M. Schuurmans
    • 1
  • V. Oldenborg
    • 1
  • A. J. van der Molen
    • 1
  • L. M. G. van Golde
    • 1
  1. 1.Laboratory of Veterinary BiochemistryState University of UtrechtUtrechtThe Netherlands

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