布地奈德对哮喘小鼠气道重塑及JAK1/STAT6表达的影响  

Budesonide attenuates airway remodeling and modules the expression of Janus protein tyrosin kinase 1,and signal transducers and activators of transcription 6 in asthma:an experiment with mice

作　　者：陈晓红[1] 钟南山[1] 张卫东[1] 曹智臻[1] 何梦璋[1] 罗群[1] 任筱兰[1] 李时悦[1] Li L 

机构地区：[1]广州医学院第一附属医院,广州呼吸疾病研究所510120 [2]Centocor Inc.USA

出　　处：《中华医学杂志》2007年第23期1627-1632,共6页National Medical Journal of China

基　　金：中国博士后基金(2005038495);广东省自然基金(5301082)

摘　　要：目的研究布地奈德(BUD)对支气管哮喘小鼠气道重塑及对 JAK1、信号转导子和转录激活子6(STAT6)表达的调控作用。方法将30只雌性 BALB/c 小鼠随机分为对照组、哮喘组和BUD 组。以鸡卵蛋白为过敏原致敏和激发动物,建立慢性哮喘气道重塑模型。对照组动物采用生理盐水致敏和激发动物,BUD 组在每次用过敏原激发前2 h 鼻腔滴入 BUD(1 mg/kg)。用 BUXCO 小鼠肺功能仪测定动物气道反应性,对肺组织切片行过碘酸雪夫(PAS)和 Masson 染色,就支气管周围炎细胞浸润及杯状细胞增殖进行评分,测定肺组织羟脯氨酸含量。应用免疫组化、免疫印迹及 RT-PCR方法分别检测 JAK1、STAT6、α-SMA 的蛋白及α-SMA mRNA 表达的变化。结果哮喘组动物气道反应性明显高于对照组,logPC100分别为0.62±0.78和1.88±0.34,P<0.01。BUD 组小鼠气道反应性低于哮喘组(1ogPC100为1.79±0.18,P<0.01)。与哮喘组相比,BUD 组气道上皮增生得到改善,上皮下胶原沉积减少,α-SMA 蛋白和 mRNA 表达也较哮喘组低。BUD 组黏液指数计分为(1.35±0.26)分,肺羟脯氨酸含量为(284±16)μg/100 mg,BALF 中 IL-4和 IL-13含量为(7.3±0.6)ng/L、(10.6±0.9)ng/L,与哮喘组比较差异有统计学意义(P<0.01)。STAT6主要表达于气道上皮,分布于细胞胞质、胞核。与对照组相比,哮喘组和 BUD 组的 STAT6分布未见明显变化。免疫印迹检测显示哮喘组的 JAK1和 STAT6蛋白表达均强于对照组,BUD 则能明显下调这两种信号蛋白的表达。相关分析表明,JAK1和 STAT6含量与气道 PAS 分数、α-SMA 蛋白表达及肺组织羟脯氨酸含量分别呈正相关,也与 BALF 中 IL-4和 IL-13水平呈正相关(均 P<0.05)。结论 BUD 抑制哮喘气道重塑。BUD可能通过下调 JAK1和 STAT6的表达,从而抑制了依赖于 JAK1/STAT6通路的气道重塑。Objective To investigate the effects of budesonide （BUD） on the airway remodeling and the expression of Janus protein tyrosin kinases 1 （JAK1） and signal transducer and activator of transcription 6 （STAT6） in asthma.Methods Thirty female Balb/c mice were randomly divided into 3 equal groups： control group; asthma group, sensitized on day 1,8, and 15 and challenged from day 21 to 52 with periodically repeated intranasal drip of ovalbumin （OVA）; and BUD treated group, undergoing intranasal drip of OVA as mentioned above and intranasal administration of BUD 2 hours before each OVA challenge. 24 h after the final OVA inhalation an invasive single-chamber whole body plethysmograph was used to assess the airway responsiveness. Then bronchoalveolar lavage fluid （BALF） was obtained and ELISA was used to measure the contents of interleukin （IL）-4 and IL-13. The mice were killed and their lungs taken out. HE staining and periodic acid Schiff （PAS） staining were used to observe the airway score of goblet cells. Peribronchiolar collagen deposition was imaged in Masson-stained lung sections. Biochemical assay was used to determine the total lung tissue level of collagen. Potass hydrolyse method was used to examine the content of hydroxyproline in the lung tissue. Western blotting was used to detect the protein expression of α-smooth muscle actin （SMA） , JAK1, and STAT6. RT-PCR was used to detect the mRNA expression of α-SMA. Results The value of LogPC100 of the asthma group was 1.88 ±0. 34 , significantly higher than those of the BUD and control groups （ 1.79 ± 0. 18 and 0. 82 ± 0. 78 respectively, both P = 0. 000）. The airway score of goblet cells of the asthma group was 3.05 ± 0. 23, significantly higher than those of the BUD and control groups （ 1.35 ± 0. 26 and 0. 40 ± 0. 13 respectively, both P 〈 0. 01 ）. The hydroxyproline content of the asthma group was （459 ± 47） μg/100 mg tissue , significantly higher than those of the BUD and control groups [ （ 284 ± 16 �

关 键 词：糖皮质激素类 哮喘 JAK1 信号转导子和转录激活子6 

分 类 号：R562.25[医药卫生—呼吸系统]