The Mechanism Of Sit On Alleviating Oxidative Stress And Cell Apoptosis In Asthma

objective: Cell apoptosis and oxidative stress have been reported to make a difference in the pathogenesis of allergic asthma Specific immunologic therapy is the only disease-modifying treatment for allergic individuals.Therefore we sought to determine whether house dust mite specific immunologic therapy could alleviate oxidative stess and cell apoptosis in asthma and futher mechanism research.Methods:This study is divided into animal experiment and cell experiment.Animal experiment:32 6-8 weekly Balb/C mices are randomly divided into four gourps,control group,asthma group,SIT group,anti-oxitative stress group(NAC group).Mices in Asthma group,SIT group,and NAC group are sensitized with house dust extracts.SIT group and NAC group will be treated before stimulation.And the PBS group are treated with PBS in all operations.The detection of airway hyperresponsiveness will be done within 24 h after last stimulation.Then,mices will be sacrificed.Cytokines and ROS((Reative Oxygen Species))in BALF,hematoxylin-eosin staining of lung tissue,periodic Acid-Schiff staining of lung tissue,TUNEL apoptosis experiment,ROS in lung frozen section,MDA in lung tissue number of Treg cells will be detected.Cell experiment:Three groups will be set,normal group,IL-25-low concentration group,IL-25-high concentration group.Cell will be treated with human recombined IL-25.Andcell will be collected after 12 h and 24 h.The cells will be used to do Annexin V-PI and ROS detection.Results: 1.Animal experiment:(1)Airway hyperresponsiveness: AHR, determined using Penh values, in mice in asthma group are higher than control mice.And SIT also decreased the Penh value in Der f mice(P<0.05).(2)Cell counts in BALF: the number and percentages of eosinophils,neutrophils in the BALF of nontreated Der f mices significantly are higher than those of control mices.(3)HE staining:mices in SIT and NAC group have less airway inflammatory infiltration in lung tissue compare with der f mices.(4)PAS staining:mices in SIT and NAC group have less mucus secretion in lung tissue campare with der f mices.(5)TUNEL detection:SIT and NAC group has less cell apoptosis in lung tissue compared with der f mices.(6)ELISA of cytokine,s Ig E and s Ig G1:SIT significantly inhibited the production of serum s Ig E,s Ig G1,and TH2 cytokine such as IL-4,IL-5,IL-13.And SIT also reduce the level of IL-25.(7)Detection of MDA:SIT significantly reduce the concentration of MDA in lung tissue.(8)Detection of ROS in BALF:SIT and anti-oxidative therapy reduce the production of ROS in BALF.(9)Number of Treg cells:SIT significantly increase the number of Treg cells in lymphocyte isolated from splenocyte.Cell experiment:(1)Annexin V-PI detection:IL-25 could increase the number of apoptosis in 16 HBE.(2)ROS in FCM:IL-25 could induce the production of ROS by 16 HBE. Conclutions:Animal experiment:1 SIT can alleviate the airway hyperresponsiveness,inhibite the production of s Ig E in serum,and reduce theproliferation of inflammatory cells by increase the number of Treg cells.2 SIT may reduce the production of some Th2 cytokine and the production of IL-25.3 SIT may alleviate the oxidative stress and cell apoptosis in lung tissue.Cell experiment: IL-25 could increase the number of cell with apoptosis procession and the production of ROS in 16 HBE cells.