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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(P<0.05,P<0.01);蔗糖水偏好程度、悬吊实验翻动次数显著下降(P<0.01);悬吊实验静止时间明显增加(P<0.01);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(P<0.05,P<0.01,P<0.01),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(P<0.05,P<0.01);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(P<0.01);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。
Objective To simulate the pathogenic characteristics of clinical syndrome of liver-qi stagnation syndrome in clinical bronchial asthma. To establish a rat model of psychological stress asthma by sensitization and restraint brakes of ovalbumin (OVA) mixture. Through macroscopic characterization, behavior, lung function, peripheral blood inflammatory cells, Bronchial pathology and other objective indicators for evaluation. Methods Thirty-two male SD rats were randomly divided into 4 groups: normal group, stress group, asthma group and stress asthma group. The stress group and the stress asthma group were restrained once a day at different times, on the first day for 2 hours, and every day after that for 10 minutes more than the previous day. The asthma group and the stressed asthma group were intraperitoneally injected with OVA, hydrogen Aqueous adjuvant mixture 1 m L sensitized. 2 weeks after inhalation of 1% OVA inhalation 20 min excitation, continuous 14 d. Normal group sensitized and challenged with saline, and not bound stimulus. After 28 days, macroscopic characterization, behavior, lung function, inflammatory cells in peripheral blood and histopathological changes of bronchial lung tissue were observed. Results The rats in the asthmatic group showed diminished interest, dull dull hair, loose stools, reduced food intake and slow weight gain. In the process of stimulation, there were bites of the nose and mouth, body and abdomen, cough, sneeze and throat wheezing Tone and other performance. Compared with the normal group, the scores of vertical movement in the stress asthma group decreased significantly (P <0.05, P <0.01); the degree of preference of sucrose water decreased significantly (P <0.01); the resting time of the suspension experiment increased significantly (FEV0.3) / forced vital capacity (FEV0.3), forced expiratory volume (FEV0.3), forced expiratory volume (FEV0.3) FVC decreased significantly (P <0.05, P <0.01, P <0.01), mean inhalation mean airway resistance (mean Ri) and mean expiratory airway resistance (average Re) ; The ratio of neutrophil, lymphocyte and eosinophil in peripheral blood increased (P <0.01); the infiltration of inflammatory cells in lung tissue was obvious. Compared with the stress group or the asthma group, macroscopic characterization, behavior, symptoms and signs of asthma, pulmonary function, bronchial histopathology, and the ratio of inflammatory cells in peripheral blood in asthmatic rats were significantly different. Conclusion The rat model of psychological stress asthma was established successfully by sensitization and restraint bracketing with OVA mixture, which accords with the clinical features of liver-qi stagnation syndrome in bronchial asthma and could be used in the basic research of emotional asthma.