The Inhibition Of Wnt/?-catenin On The Airway Epithelium Repairment After Injury Caused By Glucocoritiods On The Asthmatic Mice

PART1 THE ESTABLISHMENT OF OVA-INDUCED ASTHMA MODELObjective:To establish the OVA-induced asthma model.Methods:BABL/C mice fed with special desensitization fodder for 3 weeks were randomly divided into two groups:PBS and OVA, n=12. The OVA mice were injected intraperitoneally with 200 ul liquid containing 100 ?g OVA, 100?l AL (OH3) and 100?l PBS on day 0 and 14; And then 1%OVA nebulization at 21-30days.The PBS mice were only administering of 200ul PBS and nebulizated with PBS.TO evaluate the asthma model by observing the mice symptoms and testing the airway responsiveness by lung function monitoring system under the different concentrations of MCH, and the pathologic change in the mice lung was detected by HE staining, and the total cell and its'classified cells were counting in mice Bronchoalveolar Lavage Fluid(BALF).Results:Compared with the PBS group, under the stimulating of MHC, the OVA group mice appeared the symptoms including cyanosis and moving restlessly and significantly airway hyporesponsiveness. The severity of these symptoms was enhanced with the increasing of concentration of MCH. And the pathologic inflammation was observed in OVA mice lung and the total cell counts and eosinophilic cell counts were increased significantly in mice BALF.Conclusion:The OVA-induced asthma model were successfully establishedPART 2 THE MECHANISM OF Wnt-SIGNALING PATHWAY IN THE INHIBITION OF THE AIRWAY EPITHELIUM REPARATION CAUSED BY GLUCOCORTICOIDSObjective:To explore the mechanism of Wnt-signaling pathway in the process of inhibition of the airway epithelium reparation caused by glucocorticoids.Methods:In vivo experiment BABL/C mice were randomly divided into 4 groups:PBS, OVA, PBS+DEX, OVA+DEX, n=12. At 28-30 days OVA-induced asthma model, PBS+DEX group and OVA+DEX group were intraperitoneal injected 5mg/kg DEX. PBS group and OVA group were deal with PBS. The change of the airway epithelium reparation was detected by immunohistochemical test, and the expression of the molecules related with Wnt-signaling pathway including Wnt7b?LRP6?CyclinD1 and the cadherins molecules E-cadherinl in mice lung tissue were detected by Q-PCR. In vitro experiment the human bronchial epithelial cells (16 HBE) were cultured in DEX conditional medium, the expression of Wnt7b? LRP6?CyclinD1 in 16 HBE were detected by Q-PCR, and the cell cycle of 16 HBE were tested by Flow Cytometer.Results:In vivo after treatment of DEX, compared with OVA+PBS group, OVA+DEX mice had significantly lung tissue injury and low expression of E-cadherin 1 (P< 0.05),and the expression of Wnt7b?LRP6? CyclinD1 were increased significantly in lung tissue(P<0.05). In vitro experiment after cultured in DEX conditional medium, the expression of Wnt7b?LRP6?CyclinD1 were significantly increasing 16 HBE and whose cell cycle was retardant at G0/G1.Conclusion:The Wnt-signaling pathway plays an important role in the process of inhibition of the airway epithelium reparation caused by glucocorticoids.