Apoptosis induced by ischemia and reperfusion in experimental lung transplantation

Abstract

Background. Apoptosis is a distinct form of single-cell death in response to injury. Time course of apoptosis in lung parenchymal cells during posttransplant reperfusion and the influence of oxygen content during preservation on apoptosis of parenchymal cells are studied.Methods. Orthotopic syngenic single left lung transplantation was performed in male Fischer (F344) rats after 18 hours of cold ischemia (n = 5 in all groups). Apoptotic cells were stained by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique. Strictly TUNEL-positive pneumocytes were counted on anonymized slides by a pathologist on 100 fields (×400) per specimen (mean ± SEM).Results. The peak of apoptotic pneumocytes occurred 2 hours after reperfusion (16.8 ± 2.2 pneumocytes/100 fields [p/100f]; p = 0.000012 vs controls, lungs fixed after 18 hours of ischemia), whereas the lowest level of apoptotic pneumocytes was seen in lungs fixed after harvest (1.4 ± 0.51 p/100f) and lungs not undergoing reperfusion (2.8 ± 0.49 p/100f). Four hours after reperfusion, the number of apoptotic pneumocytes was lower than 2 hours after reperfusion (13.6 ± 3.1 p/100f; p = 0.00032 vs controls), with a further decline at 8 hours (6.4 ± 1.5 p/100f) and 12 hours after reperfusion (4.0 ± 1.2 p/100f). Interestingly, lungs inflated with N2 before storage revealed a significantly lower level of TUNEL-positive pneumocytes 2 hours after reperfusion (8.8 ± 2.0 p/100f) compared with lungs inflated with 100% O2 (p = 0.0052).Conclusions. Apoptosis of pneumocytes after posttransplant lung reperfusion is a very early event. Prolonged hypothermic preservation without reperfusion, however, does not lead to an elevated rate of apoptotic pneumocytes in lung grafts.