钙池操纵的钙通道在屋尘螨诱导哮喘小鼠气道上皮屏障破坏中的作用

Role of store-operated Ca^(2+)entry in house dust mites-induced dysfunction of airway epithelial barrier in asthmatic mice

目的:研究钙池操纵的钙内流(SOCE)在哮喘小鼠气道上皮屏障破坏中的作用及可能机制。方法:建立屋尘螨(HDM)致敏的C57BL/6急性过敏性哮喘小鼠模型,分为正常对照组、哮喘模型组和SOCE阻断剂BTP-2干预的哮喘模型组,剔除造模过程中死亡鼠和肺泡灌洗失败小鼠,每组10只。肺组织HE染色评估肺组织炎症,酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液(BALF)中IL-13和血清中IgE水平。右旋糖酐通透性实验评估屏障通透性,肺免疫荧光及肺组织化学染色评估连接蛋白(ZO-1和occludin)的分布和表达,RT-PCR和Western Blot评估肺组织中连接蛋白的表达。结果:与正常对照组相比,HDM致敏的小鼠气道炎症加重,肺组织ZO-1、occludin的mRNA和蛋白表达水平显著降低,同时右旋糖酐通透性增高(P<0.0001);与哮喘模型组相比,BTP-2干预组小鼠气道炎症减轻,ZO-1、occludin的mRNA和蛋白表达水平较哮喘组增高,右旋糖酐通透性亦明显下降(P<0.0001)。结论:SOCE及其介导的钙信号可能参与了HDM诱导的小鼠气道上皮屏障的破坏。

Objective:To investigate the potential role and mechanisms of store-operated Ca^(2+)entry(SOCE)-mediated calcium signaling in airway epithelial barrier dysfunction of asthmatic mice.Methods:C57BL/6 mice were divided into the control group,asthma model group,and asthma mod-el group treated with SOCE blocker BTP-2.Acute allergic asthma models were established with house dust mite(HDM)sensitization and challenge.Lung inflammation was evaluated by HE stain-ing.Enzyme-linked immunosorbent assay(ELISA)was used to assess the levels of IL-13 in bron-choalveolar lavage fluid(BALF)and IgE in serum.The airway epithelial integrity was evaluated by the FITC-dextran permeability test.The expression of ZO-1 and occludin in airway epithelial was evaluated with immunofluorescence and histochemistry,and the mRNA and protein expression of ZO-1 and occludin in lung tissue was evaluated by RT-PCR and Western Blot.Results:The airway in-flammation in asthmatic mice was aggravated as compared with the mice in control group.Compared with the control mice,the mRNA and protein levels of ZO-1 and occludin in lung tissue were signifi-cantly reduced,and the permeability of FTIC-dextran was increased in asthmatic mice(P<0.0001).BTP-2 treatment significantly reduced the airway inflammation,increased the mRNA and protein ex-pression of ZO-1 and occludin,and decreased the permeability of FTIC-dextran in asthmatic mice(P<0.0001).Conclusion:SOCE may participate in HDM-induced dysfunction of the airway epithelial barrier in asthmatic mice.