Prta00

The left lung was lavaged three times with 3 ml cold sterile saline. BALF samples were centrifuged (206 × g for 10 min, 4°C) and were stored at −80°C (28 (link)). Interleukin (IL)-6 and tumor necrosis factor-α (TNF-α) expression levels in the BALF were measured with ELISA kits (cat. nos. PR6000B and PRTA00, respectively; R&D Systems, Inc., Minneapolis, MN, USA). The upper lobe of the lung graft was desiccated at 80°C for 72 h to measure the W/D ratio. A portion of the lung graft tissue was frozen, homogenized and processed for MPO detection with a colorimetric assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) the change in spectrophotometric absorbance at 460 nm and was expressed as the optical density unit per gram of tissue.

Twelve rats per group were anesthetised intraperitoneally with pentobarbital (40 mg/kg) and the lungs were lavaged through a tracheal cannula by instillation of 25 mL/kg aliquots of 4°C sterile phosphate-buffered saline solution. Bronchoalveolar lavage fluid (BALF) was retrieved by gentle aspiration and the lavage procedure was repeated three times. All BALF samples from the same rat were pooled and immediately centrifuged at 1200 rpm for 10 min at 4°C. The cell-free supernatant was obtained and stored at −20°C for analysis of tumour necrosis factor-α (TNF-α) and interleukin-8 (IL-8) levels with an ELISA method using commercially available kits (reference nos. PRTA00 and FAB2164P-100, respectively; R&D Systems, Minneapolis, Minnesota, USA).