气道上皮细胞阴离子交换蛋白2在豚鼠支气管哮喘模型中表达的实验研究
Experimental research on expression of anion exchanger 2 in airway epithelial cells of guinea pig model of allergic brochial asthma
摘要目的 研究支气管哮喘(简称哮喘)豚鼠气道上皮细胞阴离子交换蛋白2( anion exchanger2,AE2)的表达及意义。方法取20只豚鼠随机分为2组,每组用卵清白蛋白(OVA)致敏,然后分别用生理盐水(对照组)和OVA(哮喘组)雾化吸入激发豚鼠哮喘。在连续雾化3d后,刮取气道上皮以免疫印迹( western blotting)和逆转录-聚合酶链反应(RT-PCR)法作AE2蛋白及其mRNA表达水平的检测。结果①呼吸频率:激发后对照组与哮喘组比较差异有统计学意义(P<0.01);同组内激发前后比较对照组内差异无统计学意义(P>0.05),哮喘组内比较差异有统计学意义(P<0.01)。②血清IgE浓度和血嗜酸粒细胞(EOS)计数:对照组IgE浓度和EOS计数与哮喘组比较,差异有统计学意义(P<0.01)。③豚鼠肺组织病理形态改变:光学显微镜观察发现:对照组,豚鼠肺组织未见病理形态改变,哮喘组则有明显哮喘病理形态改变。④AE2蛋白及其mRNA的表达:结果表明对照组豚鼠气道上皮有较高的AE2蛋白和mRNA表达量,哮喘组相对对照组表达明显降低,差异有统计学意义(P<0.01)。结论 AE2蛋白及mRNA在正常豚鼠气道上皮有一定的基础表达。过敏性哮喘模型豚鼠气道上皮细胞表达的AE2蛋白及mRNA表达明显减少。
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abstractsObjective To the study expression and significance of AE2 protein and mRNA inepithelium of asthmatic guinea pigs. Methods Twenty guinea pigs were randomly divided into control group and asthma group, with 10 guinea pigs per group. The asthmatic model of guinea pigs were established by immunization with intramuscular injected ovalbumin(OVA). The bronchial provocation test by OVA was used in each group,and asthma models were made after consecutive 3 d challenge. To scrape the epithelium in airway of guinea pig and examine the levels of AE2 protein and mRNA by western blotting and RT-PCR technique. Results ① The detection of respiratory frequency: the control group relatively had a significant difference ( P <0.01) with the asthma group after challenge and had no significant difference ( P >0.05) in the same control group, the asthma group relatively has a significant difference ( P <0.01). ②The detection of serum IgE and count eosinophil of blood:compared with the concentration and count of IgE of the control group, the asthma group had significant difference ( P <0.01). ③Pulmonary tissue pathologically morphological change:the control group, guinea pig lung tissue slices had no pathologically morphological change. But the asthma group had a distinct picture. ④The levels of AE2 protein and mRNA:the AE2 protein and mRNA appeared a basic expression in the control group. AE2 protein and mRNA expression of the asthma group decreased obviously compared with the control group ( P < 0.001 ). Conclusions AE2 protein and mRNA appeared their basic expression in normal guinea pigs. But expression of AE2 protein and mRNA in asthma group downregulated.
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