Light microscopy

For histological analysis, insert the ophthalmic scissors vertically and gently into the nasal cavities on both sides, cut the nasal back, lift the nasal bone upward, expose the nasal septum and nasal cavity of mice, take the nasal septum mucosa and fix it in 10% neutral formalin, and then place the mice in 75% alcohol to fully infiltrate and disinfect, fix the mice with a syringe needle and foam plate, extract the lung, fully strip the surrounding fat, blood vessels, bronchus, and connective tissue, and fix it in the fixative solution. Fixed nasal and lung tissues were embedded in paraffin and tissue sections, 4 mm thick, were fixed to microscope slides and deparaffinized. It was stained with hematoxylin–eosin (H&E) and then analyzed under light microscopy (HITACHI, Japan). Nasal and lung histopathological changes were analyzed.

The following reagents were used: CBD (Sigma, United States); colchicine (Beyotime, China); CCl₄ (Aladdin, China); peanut oil (Yuanye, China); ELISA kits (Novus, United States); hematoxylin and eosin (HE) and Masson assay kits (Beyotime, China); aspartate aminotransferase (AST) and hyaluronic acid (HA) kits (Nanjing Jiancheng Bioengineering Institute, China); RIPA lysis buffer and a BCA kit (Solaibao, China); COX-2, p-IκBα, and IκBα antibodies (Wanleibio, China); p-NF-κB, NF-κB, p-p38 MAPK, and p-38 MAPK antibodies (CST, United States); TGF-β1, α-SMA, COL-I, PPAR-α and GAPDH antibodies (Abcam, United States); horseradish peroxidase-labeled secondary antibodies (Bioprimacy, China); chemiluminescence (ECL) color developing solution (Merck, United States); an RNeasy mini kit (Axygen, United States); a PrimeScript RT reagent kit with gDNA Eraser and SYBR Green Master Mix (Takara, Japan); an automatic chemical analyzer (Hitachi, Japan); and light microscopy (Nikon, Japan).