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Reduced vascular endothelial growth factor correlates with alveolar epithelial damage after experimental ischemia and
Antonia Fehrenbach1, Thomas Pufe, Thorsten Wittwer
1Division of Electron Microscopy, Center of Anatomy, University of Göttingen, Göttingen, Germany. afehrenb@med.uni-marburg.de
Background:
After clinical lung transplantation, the amount of vascular endothelial growth factor (VEGF) was found to be decreased in the bronchoalveolar lavage from lungs with acute lung injury. Since Type II pneumocytes are a major site of VEGF synthesis, VEGF depression may be an indicator of pulmonary epithelial damage after ischemia and reperfusion.
Methods:
Using an established rat lung model, we investigated the relationship between VEGF protein expression, oxygenation capacity and structural integrity after extracorporeal ischemia and reperfusion (ischemia 6 hours at 10 degrees C, reperfusion 50 minutes) and preservation with either low-potassium dextran solution (Perfadex 40 kD, n = 8) or Celsior (n = 6). Untreated, non-ischemic lungs served as controls (n = 5 per group). Perfusate oxygenation was recorded during reperfusion. An enzyme-linked immunoassay (ELISA) for VEGF protein and reverse transcription-polymerase chain reaction (RT-PCR) for mRNA splice variants were determined on tissue collected from the left lungs, whereas the right lungs were fixed by vascular perfusion for VEGF immunohistochemistry as well as structural analysis by light and electron microscopy. Tissue collection by systematic uniform random sampling was representative for the whole organ and allowed for quantification of structures by stereological means.
Results:
After ischemia and reperfusion, the 3 major VEGF isoforms, VEGF(120), VEGF(164) and VEGF(188), were present. VEGF protein expression was reduced, which correlated significantly with perfusate oxygenation (r = 0.736; p = 0.002) at the end of reperfusion. It was inversely related to Type II cell volume (r = 0.600; p = 0.047). VEGF protein was localized by immunohistochemistry in Type II pneumocytes, alveolar macrophages as well as bronchial epithelium, and staining intensity of Type II cells was reduced after ischemia and reperfusion. Alveolar edema did not occur but significant interstitial edema accumulated around vessels and in the blood-gas barrier, which showed a higher degree of epithelial damage after preservation with Celsior compared with the other groups.
Conclusions:
Depression in VEGF protein expression can be considered an indicator for increased alveolar epithelial damage. Preservation with low-potassium dextran solution resulted in improved oxygenation and tissue integrity compared with Celsior.