c—Jun氨基末端激酶磷酸化在支气管哮喘大鼠气道重塑中的作用及糖皮质激素的影响  

The role of phosphorylation of c-Jun NH2-terminai kinase in airway remodeling of asthmatic rats and the effect of glucocorticoids

作　　者：林立[1] 管小俊[1] 李昌崇[1] 苏苗赏[1] 张维溪[1] 叶乐平[1] 王强[1] 陈小芳[2] 

机构地区：[1]温州医学院附属育英儿童医院呼吸科浙江省小儿呼吸诊疗研究中心,325027 [2]温州医学院附属育英儿童医院儿科研究所,325027

出　　处：《中华结核和呼吸杂志》2010年第3期188-192,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金资助项目（30271383）

摘　　要：目的研究c—Jun氨基末端激酶（JNK）磷酸化在支气管哮喘（简称哮喘）气道重塑中的作用，探讨糖皮质激素对白细胞介素（IL）-1β、JNK及哮喘气道重塑的影响。方法将48只SD大鼠按随机数字表法分为对照组、哮喘组、布地奈德组和地塞米松组，每组12只，实验组以卵清白蛋白致敏和激发复制哮喘气道重塑模型，干预组分别于每次雾化激发前以布地奈德雾化或地塞米松腹腔注射干预，对照组以生理盐水代替卵清白蛋白致敏和激发。采用图像分析技术测定支气管壁厚度（Wat）和平滑肌厚度（Wam），ELISA法测定血清、BALF中IL-1β浓度，免疫组织化学检测肺内磷酸化JNK（P-JNK）及其下游物磷酸化c—Jun蛋白表达，Westernblot检测肺匀浆P—JNK表达，直线相关分析Wat、Warn与P—JNK蛋白的平均吸光度（mA）的相关性以及P—JNK蛋白的mA与血清、BALFIL-1β浓度的相关性。结果哮喘组气道壁厚度较对照组明显增加，其血清和BALF中IL-1β浓度[分别为（81±4）ng／L、（331±15）ng／L]高于对照组[（53±6）ng／L、（130±9）ng／L]（t值分别为-8．62、-24．10，均P〈0．01）；免疫组织化学显示哮喘组P-JNK和P-c—Jun蛋白表达增高；Westernblot检测哮喘组P—JNK蛋白的mA为1．66±0．16高于对照组的1．00±0．00（t=-8．35，P〈0．01）；布地奈德、地塞米松均可抑制JNK的磷酸化；各组Wat、Wam与P—JNKmA均呈高度正相关（r值分别为0．700、0．769，均P〈0．01，n=48），P-JNK的mA与血清、BALFIL—1β浓度均呈显著正相关（r值分别为0．689、0．805，均P〈0．01）。结论JNK磷酸化与哮喘气道重塑密切相关；糖皮质激素能抑制JNK磷酸化，其机制之一可能是下调IL-1β表达。Objective To study the role of phosphorylation of c-Jun NH2-terminal kinase （JNK） in asthmatic airway remodeling and to explore the effect of glueoeorticoids on IL-1β, JNK and airway remodeling. Methods Forty-eight 445 week old male SD rats were randomly divided into 4 groups with 12 rats in each group： the control group, the asthma group, the budesonide （BUD） group, and the dexamethasone（DXM） group. The asthma airway remodeling models were made by intra-peritoneal injection of ovalbumin（OVA） on days 1 and 8 and inhalation of OVA every other day for 12 weeks since day 15. The BUD group undenvent inhalation of BUD 30 rain before every inhalation; the DXM group received intraperitoneal injection of DXM 30 rain before every inhalation; while the control group received normal saline instead of OVA. The histopathology and ultrastructural changes of pulmonary tissues were observed by light microscope and transmission electron microscope（TEM）. The total bronchial wall thickness （Wat） and the airway smooth muscle thickness（Warn） were measured by image analysis system. The concentrations of IL-1β in serum and BALF were tested by sandwich ELISA. The protein expressions of P-JNK and P-e-Jun were detected by immunohistoehemistry technique. Lung tissue extracts were analyzed for phosphorylation of JNK by Western blot. Linear correlation analysis was used to evaluate correlations between V/at and P-JNK protein（mA）, Wam and P-JNK protein（mA）, P-JNK protein （mA） and levels of IL-1βin serum, P-JNK protein （mA） and levels of IL-1β in BALF. Results In the asthma group, HE-staining showed inflammatory cell infiltration around bronchi and mucous gland hyperplasia. TEM examination showed airway smooth muscle and collagen fiber proliferation, and widening of intercellular distance. The Wat and Wam of the asthma group were significantly higher than those of the control group, while the thickness of airway wall in the glucocorticoid intervention groups became significantly decreased. The

关 键 词：哮喘 JNK丝裂原活化蛋白激酶类 白细胞介素1Β 糖皮质激素类 气道重塑 

分 类 号：R562.25[医药卫生—呼吸系统]