急性支气管哮喘小鼠肺组织水通道蛋白1表达的变化及乙酰唑胺对水通道蛋白1表达的影响  

Effect of acazolamide on expression of aquaporin 1 in lung tissue of acute allergic bronchial asthma mice

作　　者：唐顺广[1] 周瑞祥[1] 贲素琴[2] 倪松石[2] 

机构地区：[1]高邮市人民医院呼吸内科,225601 [2]南通大学附属医院呼吸内科,226001

出　　处：《中华哮喘杂志（电子版）》2009年第5期6-14,共9页Chinese Journal of Asthma(Electronic Version)

摘　　要：目的通过水通道蛋白1(aquaporins1,AQP1)在急性过敏性支气管哮喘(简称哮喘)小鼠肺组织表达的变化,以及乙酰唑胺对AQP1表达的影响,探讨哮喘的分子发病机制,为哮喘防治提供新思路。方法40只C57BL/6小鼠随机分为A、B、C、D、E5组,其中A组为磷酸盐缓冲液(phosphate buffer saline,PBS)非哮喘组、其余4组建立哮喘模型,B组、C组、D组为乙酰唑胺干预组(300mg/kg·200mg/kg100mg/kg)和哮喘对照组(E组),干预组在激发阶段给予不同剂量的乙酰唑胺腹腔注射。5d后处死小鼠观察肺组织形态学改变;测肺湿干重比;支气管肺泡灌洗液的细胞总数、分类计数以及白介素5(IL-5)和γ干扰素(IFN-γ)水平;用RT-PCR、Western blot和免疫组织化学及免疫荧光法检测肺组织AQP1mRNA及AQP1的水平和分布情况。结果哮喘对照组肺组织的湿干重比值比非哮喘组明显增高(P<0.05);乙酰唑胺干预组肺组织的湿干重比值较哮喘对照组显著减低(P<0.05),与剂量呈负相关;②哮喘对照组BALF中细胞总数、嗜酸粒细胞数和IL-5水平均高于非哮喘组(P<0.05),IFN-γ水平低于非哮喘组(P<0.05),治疗后细胞总数、嗜酸粒细胞数中性粒细胞淋巴细胞均明显下降(P<0.05),且与剂量呈正相关;③哮喘对照组AQP1在气管黏膜上皮、微血管内膜上皮、支气管周围血管床、炎症细胞呈强阳性表达,其表达水平明显高于非哮喘组(P<0.01),使用乙酰唑胺干预后表达明显低于哮喘对照组(P<0.05),下降的幅度与剂量呈正相关性;但AQP1mRNA检测结果无明显差异(P>0.05);④哮喘对照组肺组织可见较弥漫的炎性改变;而乙酰唑胺干预组炎性改变则明显减轻。结论AQP1在急性哮喘小鼠肺组织内及炎症细胞上过度表达,哮喘组呈强阳性表达;②乙酰唑胺对AQP1有显著的抑制作用,可明显改善小鼠肺部的炎性细胞浸润等病理改变,且呈剂量依赖性;③AQP1蛋白质在各组中表达差异较大,但AQP1mRNA差异�Objective To study the expression of aquaporin-1 （AQP1） in pulmonary tissues of acute allergic bronchial asthma（asthma） mice and the effects of acetazolamide （Acetazolamide, AZ） on which expression in order to uncover the mechanism of asthma and provide new view for the prevention of asthma. Methods 40 C57BL/6 mice were randomly divided into A,B,C,D,E group,mice of group A treated with phosphate buffer （Phosphate buffer saline, PBS） as non asthmatic group, and the remaining groups were established the asthma model. Group B,C and D were treated with acetazolamide （300 mg/kg, 200 mg/kg, 100 mg/kg） as intervention. Five days later the mice were killed to observe the morphological changes of lung tissue,and to measure the proportion of wet and dry; bronchoalveolar lavage fluid were collected to observe cells of the total count and IL-5 and IFN-γ levels;RT - PCR, Western blot,immunohistochemical and immunofluorescence were used to detect the level of AQPlmRNA and the distribution of AQP1 in lung tissue. Results ①Wet/dry weight ratio of lung tissues in asthma control group was significantly higher than non-asthma group（ P 〈0.05）, however, which was lower in intervention groups than that of asthma control group;②The total number of cells in BALF and the number of eosinophils and IL-5 in asthma control group were higher than non-asthmatic group （ P 〈0. 05） ,and IFN-γ level was lower in asthma control group than non-asthmatic group （P 〈0.05）. After treatment the total number of cells, the number of eosinophils,neutrophils and lymphocytes were significantly decreased （ P 〈0.05）, which were positively correlated with the dose of acetazolamide;③AQP1 were expressed in tracheal epithelium, microvaseutar endothelial cell and bronchial peripheral vascular bed, and the expression in asthma control group was significantly higher than non-asthmatic group （ P 〈0.01）, which was significantly decreased after the intervention of acetazolamide than that of asthma control group �

关 键 词：过敏性哮喘 水通道蛋白1 乙酰唑胺 白介素5 Γ干扰素 

分 类 号：R562.25[医药卫生—呼吸系统] R563[医药卫生—内科学]