Mice were euthanized, the thoracic cavity was opened, and the right main bronchus was ligated at the left and right bronchial bifurcations. Pre-cooled PBS was used to perform bronchoalveolar lavage of the right lung three times; each time, 0.4 ml BALF was recovered. The BALF was transferred to a tube, centrifuged at 1,800 × g at 4°C for 15 min and the supernatant was collected. The levels of inflammatory factors, IL-1β (cat. no. MLB00C), IL-6 (cat. no. M6000B), IL-18 (cat. no. 7625) and TNF-α (cat. no. MTA00B), in the BALF were detected by ELISA according to manufacturer's protocols. The ELISA kits were purchased from R&D Systems, Inc.. The concentration of Protein in the BALF was measured using the BCA method (Bio-Rad Laboratories, Inc.).

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