Optimal duration of ex vivo lung perfusion for heat stress-mediated therapeutic reconditioning of damaged rat donor lungs.

OBJECTIVES

Transient heat stress (HS) application during experimental ex vivo lung perfusion (EVLP) of warm ischaemic (WI) rat lungs produces a range of therapeutic benefits. Here, we explored whether different EVLP durations after HS application would influence its therapeutic effects.

METHODS

In protocol 1, WI rat lungs were exposed to HS (41.5°C, 60-90 min EVLP), and EVLP was maintained for 3, 4.5 or 6 h (n = 5/group), followed by physiological measurements (compliance, oedema, oxygenation capacity). In protocol 2, WI rat lungs treated with (HS groups) or without HS (control groups) were maintained for 3 or 4.5 h EVLP (n = 5/group), followed by physiological evaluation and measurements (lung tissue) of heat shock proteins (HSP70, HSP27, HS90, GRP78), endogenous proteins (surfactant protein-D, CC16, platelet endothelial cell adhesion molecule-1), anti-apoptotic (Bcl2, Bcl-xL) and pro-apoptotic proteins (Bcl2-associated X protein, CCAAT/enhancer binding-protein homologous protein), antioxidant enzymes (heme-oxygenase-1, nicotinamide di-phospho-nucleotide dehydrogenase quinone-1) and nitrotyrosine (oxidative stress biomarker).

RESULTS

In protocol 1, physiological variables were stable after 3 and 4.5 h but deteriorated after 6 h. In protocol 2, at 3 h EVLP, HS-treated lungs differed from controls by higher expression of HSP70 and heme-oxygenase-1, and lower CC16 expression. In contrast, at 4.5 h EVLP, HS-treated lungs displayed improved physiology, higher levels of all HSPs, preserved or increased expression of surfactant protein-D, CC-16 and platelet endothelial cell adhesion molecule-1, increased antioxidant and anti-apoptotic proteins, and reduced pro-apoptotic proteins and nitrotyrosine.

CONCLUSIONS

The protective effects of HS application during EVLP of WI-damaged rat lungs strictly depend on the duration of post-HS recovery. An EVLP duration of 4.5 h appears to optimize the therapeutic potential of HS, while maintaining lungs in a stable physiological state.