Skip to main content
PMC home page
Thorax logoLink to Thorax
. 1997 Aug;52(8):723–730. doi: 10.1136/thx.52.8.723

Lung surfactant in a cystic fibrosis animal model: increased alveolar phospholipid pool size without altered composition and surface tension function in cftrm1HGU/m1HGU mice

W Bernhard, J Y Wang, T Tschernig, B Tummler, H J Hedrich, H von der Hardt
PMCID: PMC1758618  PMID: 9337833

Abstract

BACKGROUND: Progressive pulmonary dysfunction is a characteristic symptom of cystic fibrosis (CF) and is associated with functional impairment and biochemical alterations of surfactant phospholipids in the airways. However, the fundamental question of whether surfactant alterations in the CF lung are secondary to the pulmonary damage or are present before initiation of chronic infection and inflammation has yet to be resolved in patients with cystic fibrosis but can now be addressed in CF mice that exhibit the basic defect in the airways. A study was therefore undertaken to investigate the pool sizes, composition, and function of lung surfactant in the non-infected cftrm1HGU/m1HGU mouse. METHODS: The amount and composition of phospholipid classes and phosphatidylcholine molecular species were determined in bronchoalveolar lavage (BAL) fluid and lavaged lungs by high performance liquid chromatography (HPLC). Surfactant protein A (SP- A) levels in BAL fluid were determined by ELISA and surfactant for functional measurements was isolated from BAL fluid by differential ultracentrifugation. Equilibrium and minimal surface tension of surfactant was assessed by the pulsating bubble surfactometer technique. MF1, BALB/c, C57/BL6, and C3H/He mice served as controls. RESULTS: BAL fluid of cftrm1HGU/m1HGU mice contained 1.02 (95% confidence interval (CI) 0.89 to 1.16) mumol phospholipid and 259 (239 to 279) ng SP-A. BAL fluid of MF1, BALB/c, C57BL/6, and C3H/He mice contained 0.69 (0.63 to 0.75), 0.50 (0.42 to 0.57), 0.52 (0.40 to 0.64), and 0.45 (0.27 to 0.63) mumol phospholipid, respectively. After correction for the different body weights of mouse strains, phospholipid levels in BAL fluid of cftrm1HGU/m1HGU mice were increased by 64 (52 to 76)%, 60 (39 to 89)%, 72 (45 to 113)%, and 92 (49 to 163)%, respectively, compared with controls. The amount of SP-A in BAL fluid and the composition of phospholipid as well as phosphatidylcholine molecular species in BAL fluid and lung tissue was unchanged in cftrm1HGU/m1HGU mice compared with controls. The increase in phospholipids in BAL fluid of cftrm1HGU/m1HGU mice resulted from an increased fraction of large aggregates which exhibited normal surface tension function. CONCLUSION: In cftrm1HGU/m1HGU mice surfactant homeostasis is perturbed by an increased phospholipid pool in the alveolar compartment. 




Full Text

The Full Text of this article is available as a PDF (217.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bernhard W., Linck M., Creutzburg H., Postle A. D., Arning A., Martin-Carrera I., Sewing K. F. High-performance liquid chromatographic analysis of phospholipids from different sources with combined fluorescence and ultraviolet detection. Anal Biochem. 1994 Jul;220(1):172–180. doi: 10.1006/abio.1994.1315. [DOI] [PubMed] [Google Scholar]
  4. Chace K. V., Leahy D. S., Martin R., Carubelli R., Flux M., Sachdev G. P. Respiratory mucous secretions in patients with cystic fibrosis: relationship between levels of highly sulfated mucin component and severity of the disease. Clin Chim Acta. 1983 Aug 15;132(2):143–155. doi: 10.1016/0009-8981(83)90242-5. [DOI] [PubMed] [Google Scholar]
  5. Dorin J. R., Dickinson P., Alton E. W., Smith S. N., Geddes D. M., Stevenson B. J., Kimber W. L., Fleming S., Clarke A. R., Hooper M. L. Cystic fibrosis in the mouse by targeted insertional mutagenesis. Nature. 1992 Sep 17;359(6392):211–215. doi: 10.1038/359211a0. [DOI] [PubMed] [Google Scholar]
  6. Dorin J. R., Stevenson B. J., Fleming S., Alton E. W., Dickinson P., Porteous D. J. Long-term survival of the exon 10 insertional cystic fibrosis mutant mouse is a consequence of low level residual wild-type Cftr gene expression. Mamm Genome. 1994 Aug;5(8):465–472. doi: 10.1007/BF00369314. [DOI] [PubMed] [Google Scholar]
  7. Doyle I. R., Barr H. A., Davidson K. G., Nicholas T. E. Differential changes in SP-A and disaturated phospholipids in the isolated perfused rat lung and in vivo. Am J Physiol. 1996 Sep;271(3 Pt 1):L374–L382. doi: 10.1152/ajplung.1996.271.3.L374. [DOI] [PubMed] [Google Scholar]
  8. Engelhardt J. F., Zepeda M., Cohn J. A., Yankaskas J. R., Wilson J. M. Expression of the cystic fibrosis gene in adult human lung. J Clin Invest. 1994 Feb;93(2):737–749. doi: 10.1172/JCI117028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Enhorning G. Pulsating bubble technique for evaluating pulmonary surfactant. J Appl Physiol Respir Environ Exerc Physiol. 1977 Aug;43(2):198–203. doi: 10.1152/jappl.1977.43.2.198. [DOI] [PubMed] [Google Scholar]
  10. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  11. Galabert C., Jacquot J., Zahm J. M., Puchelle E. Relationships between the lipid content and the rheological properties of airway secretions in cystic fibrosis. Clin Chim Acta. 1987 Apr 30;164(2):139–149. doi: 10.1016/0009-8981(87)90065-9. [DOI] [PubMed] [Google Scholar]
  12. Hamm H., Fabel H., Bartsch W. The surfactant system of the adult lung: physiology and clinical perspectives. Clin Investig. 1992 Aug;70(8):637–657. doi: 10.1007/BF00180279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Lewis J. F., Jobe A. H. Surfactant and the adult respiratory distress syndrome. Am Rev Respir Dis. 1993 Jan;147(1):218–233. doi: 10.1164/ajrccm/147.1.218. [DOI] [PubMed] [Google Scholar]
  15. Marino C. R., Gorelick F. S. Scientific advances in cystic fibrosis. Gastroenterology. 1992 Aug;103(2):681–693. doi: 10.1016/0016-5085(92)90866-w. [DOI] [PubMed] [Google Scholar]
  16. McCray P. B., Jr, Wohlford-Lenane C. L., Snyder J. M. Localization of cystic fibrosis transmembrane conductance regulator mRNA in human fetal lung tissue by in situ hybridization. J Clin Invest. 1992 Aug;90(2):619–625. doi: 10.1172/JCI115901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oulton M., MacDonald J., Janigan D. T., Faulkner G. T. Mouse alveolar surfactant: characterization of subtypes prepared by differential centrifugation. Lipids. 1993 Aug;28(8):715–720. doi: 10.1007/BF02535992. [DOI] [PubMed] [Google Scholar]
  18. Sugahara K., Iyama K., Sano K., Morioka T. Differential expressions of surfactant protein SP-A, SP-B, and SP-C mRNAs in rats with streptozotocin-induced diabetes demonstrated by in situ hybridization. Am J Respir Cell Mol Biol. 1994 Oct;11(4):397–404. doi: 10.1165/ajrcmb.11.4.7917308. [DOI] [PubMed] [Google Scholar]
  19. Ulane M. M., Butler J. D., Peri A., Miele L., Ulane R. E., Hubbard V. S. Cystic fibrosis and phosphatidylcholine biosynthesis. Clin Chim Acta. 1994 Oct 31;230(2):109–116. doi: 10.1016/0009-8981(94)90263-1. [DOI] [PubMed] [Google Scholar]
  20. Wang J. Y., Yeh T. F., Lin Y. C., Miyamura K., Holmskov U., Reid K. B. Measurement of pulmonary status and surfactant protein levels during dexamethasone treatment of neonatal respiratory distress syndrome. Thorax. 1996 Sep;51(9):907–913. doi: 10.1136/thx.51.9.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Golde L. M. Potential role of surfactant proteins A and D in innate lung defense against pathogens. Biol Neonate. 1995;67 (Suppl 1):2–17. doi: 10.1159/000244202. [DOI] [PubMed] [Google Scholar]

Articles from Thorax are provided here courtesy of BMJ Publishing Group

RESOURCES