Inhibition Of Airway Remodeling In Mice With Bronchial Asthma By Dabigatran Etexilateon And The Mechanistic Investigation

Background:Bronchial asthma(asthma)is a common and serious influence quality of life of patients with chronic inflammatory airway disease,the pathology is characterized by chronic airway inflammation and airway hyperresponsiveness?airway remodeling.airway remodeling in varying degrees throughout the course of bronchial asthma,is the main cause of irreversible pulmonary ventilation function obstacle.airway remodeling including hypertrophy of goblet cell hyperplasia?vascular proliferation?airway smooth muscle thickening and the extracellular matrix deposition.In recent years,studies have shown that thrombin has a proinflammatory?promote proliferation and promote the smooth muscle cell secretion of extracellular matrix,and so on,and participate in asthmatic airway remodeling.dabigatran etexilateon is a new direct thrombin inhibitor,is widely applied in anticoagulant therapy.Research has shown that dabigatran etexilateon can inhibit the secretion of extracellular matrix induced by thrombin in vitro.In vivo,dabigatran etexilateon whether inhibit airway remodeling of asthma is unclear.YAP/TAZ is a downstream effector molecule of Hippo signaling pathway,which mediates the proliferation,migration and differentiation of cells,etc.YAP/TAZ is widely expressed in lung structural cells,and is involved in the development of lung and the occurrence and development of a variety of lung diseases.Whether YAP/TAZ is involved in airway remodeling in asthma is unclear.Objective:In the mouse model of asthma,to determine whether dabigatran etexilateon can reduce airway remodeling in asthrma;to determine whether inhibition of YAP/TAZ can alleviate airway remodeling in asthma;Methods:Male BABLb/c mice aged 6-8 weeks with SPF were selected.All mice were kept in SPF grade animal chamber,sterile water and radioactive feed were fed to mice,and light/dark cycle was given for 12 hours.The mice were randomly divided into six groups:(1)blank group;(2)OVA asthma model group(OVA group);(3)OVA/dexamethasone group(the OVA/Dex group);(4)OVA/dabigatran etexiteon 5mg group(the OVA/Dab5mg group);(5)OVA/dabigatran etexiteon 5mg group(the OVA/Dab10mg group);(6)OVA/Verteporfin group(OVA/Ver group),6 mice in each group,OVA model group,OVA/Dex group,OVA/Dab5mg group,OVA/Dab10mg group and OVA/Ver group were respectively sensitized by intraperitoneal injection of sensitizing solution on day 1 and day 7.The nasal drip was then stimulated with 2mg/ml OVA solution on day 14?15?16?21?22?23,respectively.The dabigatran-etexilateon intervention group was fed with the corresponding concentration of feed on the 13th day,and the corresponding amount of solution was injected into the peritoneum of the dexamethasone and vetipofen intervention groups 30 minutes before the excitation of nasal drops.Mice were sacrificed within 24 hours after the last stimulation,and alveolar lavage fluid and lung tissue were collected to observe the pathological changes of the lung.Western blotting was used to detect t-YAP and p-YAP.Results:(1)BALF cell count results showed that BALF cells were significantly increased in the OVA group,which was statistically different from the blank group(P<0.001).Compared with the OVA group,the cell count in BALF was significantly decreased in the OVA/Dex group,and the difference was statistically significant(P<0.001).The number of cells in BALF in the OVA/dabigatran etexilateon 5mg group was significantly lower than that in the OVA group,and the difference was statistically significant(P<0.001).The number of BALF cells in the OVA/dabigatran etexilateon 10mg group and OVA/verteporfin intervention group was significantly lower than that in the OVA group,and the difference was statistically significant(P<0.05).(2)The results of HE staining showed that normal small airway and alveolar structures could be seen in the blank group,and inflammatory cells in the airway and beside blood vessels were significantly increased in the OVA group compared with the blank group(p<0.05),while inflammatory cells were significantly decreased in the dexamethasone group compared with the OVA group(p<0.05).Compared with the OVA group,the inflammatory cells in the airway and beside blood vessels were significantly reduced in the dabigatran etexilateon 5mg group(p<0.05),and the inflammatory cells were decreased in the dabigatran etexilateon 10mg group.Compared with the OVA group,the number of inflammatory cells in the airway and beside blood vessels was significantly reduced in the YAP inhibitor verteporfin group(p<0.05),which was similar to that in the dabigatran etexilateon group.(3)PAS staining results showed that the number of goblet cells in the OVA group was significantly higher than that in the blank group(p<0.001).Compared with the OVA model group,goblet cells were significantly reduced in the dabigatran etexilateon 5mg group and the YAP inhibitor verteporfin group(p<0.05),and hypertrophy was similar in the two groups.(4)The results of Masson staining showed that:compared with the blank group,the deposition of collagen in the airway and collateral vessels was significantly increased in the OVA group(p<0.05).Compared with the OVA group,the bronchial and paravascular collagen deposition was less in the dabigatran etexilateon 5mg group(p<0.05).Compared with the OVA model group,the peribronchogenic collagen deposition was less in the YAP inhibitor verteporfin group(p<0.05),which was similar to that in the dabigatran etexilateon group(5)Western blotting results showed that t-YAP expression was significantly up-regulated and p-yap expression was significantly decreased in the OVA model group,suggesting up-regulated expression of dephosphorylated YAP(activated form of YAP).Dabigatran etexilateon intervention up-regulated the expression of p-YAP,suggesting that it inhibited the expression of dephosphorylated YAP(activated form of YAP).The intervention of the YAP inhibitor verteporfin inhibited the expression of t-YAP p-yap,but the inhibition of t-YAP was more significant.Conclusion:Both dabigatran etexilateon and YAP inhibitors can alleviate airway remodeling in asthma.Dabigatran etexilateon may reduce airway remodeling in asthma by inhibiting YAP activation.