心理应激哮喘大鼠模型的建立与评价

目的模拟临床支气管哮喘肝气郁结证致病特点,运用卵清蛋白(OVA)混合液致敏和束缚制动建立心理应激哮喘大鼠模型,通过宏观表征、行为学、肺功能、外周血炎症细胞、支气管组织病理等客观指标进行评价。方法 32只雄性SD大鼠随机等分为4组:正常组、应激组、哮喘组、应激哮喘组。应激组及应激哮喘组每天在不同时间束缚制动1次,第1天2 h,以后每天均比前一天多10 min;哮喘组及应激哮喘组第1天腹腔注射含OVA、氢氧化铝佐剂的混合液1 m L致敏。2周后每天雾化吸入1%OVA 20 min激发,连续14 d。正常组致敏和激发均用生理盐水,且不做束缚刺激。28 d后观察各组大鼠宏观表征、行为学、肺功能、外周血炎症细胞分类、支气管肺组织病理改变情况。结果应激哮喘组大鼠表现为兴趣减退、毛发晦暗无光泽、便溏、饮食量减少、体重增加缓慢;激发过程中有抓咬口鼻、体腹部皮肤,呛咳、喷嚏及喉中哮鸣音等表现。与正常组相比,应激哮喘组垂直运动得分明显下降(P0.05,P0.01);蔗糖水偏好程度、悬吊实验翻动次数显著下降(P0.01);悬吊实验静止时间明显增加(P0.01);呼气峰流速(PEF)、吸气中期峰流速(FEF25%~75%)、用力呼气时间(FET)、0.3 s用力呼气量(FEV0.3)/用力肺活量(FVC)下降(P0.05,P0.01,P0.01),吸气平均气道阻力(平均Ri)、呼气平均气道阻力(平均Re)明显升高(P0.05,P0.01);外周血中性细胞比率、淋巴细胞比率及嗜酸粒细胞比率均升高(P0.01);肺组织中炎症细胞浸润明显。与应激组或哮喘组比较,应激哮喘组大鼠宏观表征、行为学、哮喘症状及体征、肺功能、支气管组织病理,外周血炎症细胞比率均有显著性差异。结论采用OVA混合液致敏和束缚制动成功建立了心理应激哮喘大鼠模型,符合支气管哮喘肝气郁结证临床特征,可用于情志哮喘的基础研究。

Establishment and evaluation of psychological stress asthmatic rat model

Objective To induce the psychological stress-asthmatic rat model by ovalbumin (OVA)mix-ture sensitization with restraint,which simulated the pathogenic factors of bronchial asthma with liver -qi stagnation pattern,as well as to evaluate the model using indicators of the macro characterization,behav-iors,pulmonary function,peripheral blood inflammatory cells,bronchial tissue pathomorphology.Meth-ods A total 32 male rats were randomly divided into four groups—control group,stress group,asthmatic group,and stress-asthmatic group.Rats of stress group and stress-asthmatic group were immobilized in restrained box once daily at different time,ten minutes more than the day before with the first duration of 2 hours.Rats of stress-asthmatic group and asthmatic group were sensitized by intraperitoneal injection 1 mL mixture of OVA with aluminium hydroxide on the first day,and after 13 days,they were stimulated by inhalation of 1 % OVA for 20 min for consecutive 14 days.The rats of control group were sensitized and stimulated by saline,without immobilizing .28 days later,the macro characterization,behaviors, peripheral blood eosinophils,pulmonary function,bronchial tissue pathomorphology were observed.Re-sults Stress asthmatic rats showed low mood,loss of interest,dull hair,loose stools,reducing food in-take,slowly weight gain,and scratched noses and abdominal skin,coughed,sneezed and wheezed in the throat when stimulated.Compared with control group,the points of vertical movement decreased signifi-cantly(P <0.05 or P <0.01).Preference for sucrose,the times of active moves in tail suspension test (TST)of stress-asthmatic rats obviously decreased(P <0.01),and the duration of immobility increased significantly(P <0.01 ).Pulmonary function indicators,including peak expiratory flow(PEF),forced expiratory flow (FEF 25% -75% ),forced expiratory volume in 0.3 s FEV(0.3]/forced expiration volume capacity (FVC)significantly declined(P <0.05,P <0.01,P <0.01 ),while mean inspiratory airway resistance (Ri)and mean expiratory airway resistance (Re)increased remarkably (P <0.05,P<0.01).Peripheral blood eosinophil cells increased(P <0.01),and infiltration of inflammatory cell in lung tissue was obvious.Compared with the stress group or asthmatic group,the signs and indicators of stress-asthmatic group were all much worse.Conlusion The psychological stress-asthmatic rat model, induced by VOA mixture with restraint,was proved to be successful.It is in accordance with clinical characteristics of bronchial asthma with liver-qi stagnation pattern,which can be used to study asthma caused by emotion .