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目的评价两种肾阳虚证支气管哮喘病证结合动物模型造模方法。方法将54只Wistar大鼠分为空白组、哮喘组、激素组、激素+哮喘组、手术组、手术+哮喘组,每组9只。激素组、激素+哮喘组大鼠采用氢化可的松注射液15 mg/(kg·d)皮下注射,手术组、手术+哮喘组采用切除双侧全部肾上腺复制为肾阳虚证候模型。哮喘组、激素+哮喘组、手术+哮喘组采用卵清白蛋白(OVA)致敏诱发哮喘。观察各组大鼠一般状态、行为学、肺功能及肺泡灌洗液中白细胞分类,同时检测血清下丘脑-垂体-肾上腺(HPA)轴功能指标[包括促肾上腺皮质激素释放激素(CRH)、肾上腺皮质激素释放激素(ACTH)、皮质醇(CORT)、干扰素-γ(INF-γ)、免疫球蛋白E(Ig E)]及肺组织各项炎症因子水平;观察肺组织病理改变。结果激素+哮喘组、手术+哮喘组肺泡灌洗液中白细胞、嗜酸性细胞总数升高,CRH、ACTH、CORT分泌减少;血清总Ig E、OVA特异性Ig E含量升高;肺组织内INF-γ表达分泌减少,白细胞介素4(IL-4)表达分泌增多(P<0.05或P<0.01);肺组织病理显示支气管出现大量炎症浸润、气道黏膜水肿。激素+哮喘组与手术+哮喘组比较,在肺泡灌洗液炎症细胞检测上无明显差异,后者HPA轴功能指标下降更加明显,前者大鼠肺组织的病理改变更加明显。结论两种肾阳虚证支气管哮喘动物模型均能够反映肾阳虚证支气管哮喘的发病特点,但采用切除双侧全部肾上腺方法的动物模型对HPA轴功能影响更明显。
Objective To evaluate the method of animal modeling by combining the two syndromes of bronchial asthma with syndrome of deficiency of kidney-yang. Methods Fifty - four Wistar rats were divided into blank group, asthma group, hormone group, hormone + asthma group, operation group and operation + asthma group. Rats in the hormone group and the hormone + asthma group were subcutaneously injected with hydrocortisone 15 mg / (kg · d), and the operation group and the operation + asthma group were all duplicated with adrenal resection for the model of deficiency of kidney yang. Asthma group, hormone + asthma group, surgery + asthma group induced by ovalbumin (OVA) sensitized asthma. The general state, behavior, lung function and the classification of leukocytes in BALF were observed in each group. Serum hypothalamic-pituitary-adrenal (HPA) axis function index [including Corticotropin Releasing Hormone (CRH) (ACTH, CORT, INF-γ, Ig E) and the levels of inflammatory cytokines in lung tissue. The pathological changes of lung tissue were observed. Results The total number of leukocytes and eosinophils in the alveolar lavage fluid of the hormone + asthma group and the surgery + asthmatic group was increased, while the secretion of CRH, ACTH and CORT was decreased; the serum total Ig E and OVA specific Ig E levels were increased; (P <0.05 or P <0.01). The lung histopathology showed a large number of inflammatory infiltrates in bronchus and airway mucosal edema. Hormone + asthma group and surgery + asthma group, there was no significant difference in the detection of inflammatory cells in the alveolar lavage fluid, the latter HPA axis decreased more significantly, the former pathological changes of lung tissue more obvious. Conclusion Both animal models of bronchial asthma with deficiency of kidney-yang syndrome can reflect the characteristics of bronchial asthma with deficiency of kidney-yang syndrome. However, the animal model with bilateral all adrenal gland resection has a more obvious effect on the function of HPA axis.