Neutrophil gelatinase-associated lipocalin as a potential novel biomarker for ventilator-associated lung injury

Xiao, Rui; Chen, Rongchang

Collection of BAL fluid, serum or lung tissue. Following treatment, animals were sacrificed via administration of 50 mg/kg sodium pentobarbital (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Peripheral blood was extracted by removing the eyeball. Following overnight storage at 4ËšC, serum was collected via centrifugation at 1,000 x g for 30 min at 4ËšC. Immediately post-mortem, the heart-lung block was dissected and BAL fluid was obtained using the method previously described (12, 13) . BAL fluid was centrifuged at 1,000 x g for 10 min at 4ËšC and the supernatant was collected for western blotting. For reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting of lung samples, the right lung was cut into samples (~1x1 cm) and flash-frozen in liquid nitrogen. For immunohistochemical and hematoxylin and eosin (H&E) staining, the left lung was fixed as previously described (14) . The vertical axis of each left lung was identified and the lung was cut perpendicular to this axis into 4-mm thick slices.