Expression Of HIF-1? And Wnt/?-catenin Associated Moleculars In The Canine Central Tracheal Stenosis

Objective The study aimed to construct the animal model of Central tracheal stenosis and then investigate the expression of HIF-1? and Wnt/?-catenin associated molecules in cartilagines tracheales.Methods 30 Chinese rural dogs were divided in 3 group: Negative control group,n=10,Sham group,n=10 and operation group,n=10.Nothing was done towards the Canines in Negative control group,while operation group Canines were fixed on the operating table and anestheted by Pelltobarbitalum Natricum,then in open laryngoscope down orotracheal intubation by using I.D.8.0 with balloon tracheal tubes,when the cannula tip above the Carina 4~6cm,hold a syringe to inject gas into the cuff and maintain pressure for 200 mm Hg in the capsule.Fixed the cannula and leaving 24 h.For Sham group,Canines were suffered the same intubation but without gas injection.After extubation,the Canines in operation group were euthanized on days of 1,7 and days 14,respectively.Then we observe the tracheal anatomy,tracheal stenosis and pathological changes of experimental dogs.Subsequently,we get lesions organization,andextract its total protein and RNA,then use real-time quantitative PCR,and ELISA or Western blot to detect low oxygen induced factor-1?(HIF-1 Alpha),vascular endothelial growth factor(VEGF),transforming growth factor ?1(TGF-?1),Leukocyte interleukin-1(IL-1),?-serial protein(?-catenin),T cell factor(TCF-4),Bone morphogenetic protein-2(BMP-2),matrix metalloproteinase-13(MMP-13)and matrix metalloproteinase inhibitor(TIMP-1)respectively on expression changes of days14 after the extubation.Results 1.The sham group and the operation group of 1 dogs died during intubation.The other dogs were successfully recovered after tracheal intubation and complications such as suffocation,accidental removal of tube did not occur.2.In the sham operation group,the tracheal mucosa was slightly congested on the day of extubation and 7 days later.There was slight inflammatory exudation and little change.There were a lot of infiltration of inflammatory cells in the operation group,involving the submucosal layer and mucosal surface bleeding.The cartilage degeneration,chondritis,cartilage necrosis and fibrous tissue hyperplasia occurred on the 7th day after extubation.14 days later,Large cartilage necrosis,fibrous tissue and capillary proliferation around the cartilage,granulation tissue filling.3.The levels of HIF-1?,VEGF,TGF-?1 and m RNA expression in the cartilagines tracheales of Negative control group and Sham group were very low,while the m RNA was significantly higher in airway constriction in the operation group.And the level of HIF-1? protein expression was similar with m RNA.Simultaneously,low levels of ?-catenin,TCF-4,BMP-2,MMP-13 and TIMP-1 was expressed in Negative control group,while levels of expressions of m RNA and protein of ?-catenin,TCF-4,BMP-2,MMP-13 and TIMP-1 in Canine in days 14 after extubation are significantly increased.Conclusions: 1.By endotracheal intubation and increasing the balloon inflation pressure can effectively establish the Central Airway stenosis model.2.The levels of VEGF?TGF-?1 and IL-1? and m RNA expression in the cartilagines tracheales are significantly increased.3.The levels of HIF-1???-catenin?TCF-4?BMP-2 and MMP-13 expression in the cartilagines tracheales are significantly increased.Prompt HIF-1? and Wnt/?-cateninmay contribute to airway constriction.