组胺对呼吸道上皮紧密连接表达的影响

Role of histamine on the expression of tight junctions gene in airway epithelia

目的检测组胺刺激下体外培养的原代支气管上皮细胞间紧密连接相关基因表达的改变。方法应用气-液界面培养方法体外培养正常人和哮喘患者原代支气管上皮细胞,应用组胺刺激上皮细胞96 h,分别应用实时荧光定量PCR和免疫荧光染色共聚焦显微镜检测紧密连接相关基因在m RNA及蛋白水平的表达。结果实时荧光定量PCR结果显示组胺刺激可以诱导某些紧密连接相关基因m RNA表达的上调或下调;免疫荧光染色共聚焦显微镜检查未发现组胺刺激可以诱导某些紧密连接相关蛋白发生形态学的改变。结论组胺可以调节人原代支气管上皮细胞紧密连接相关基因m RNA表达,有可能是慢性呼吸道炎性疾病的发病机制之一。

OBJECTIVE To investigate and elucidate the role of histamine on the regulation of tight junctions(TJs) gene expression of the airway epithelial cells.METHODS We cultured primary human bronchial epithelial cells from healthy and asthmatic individuals in the presence of histamine for 96 hours in air-liquid interface(ALI) culture system.The TJs expression on the level of mRNA and protein was determined by RT-qPCR and immunofluorescence(confocal microscopy),respectively.RESULTS A RT-qPCR analysis revealed that the mRNA expression of several TJs were up- or down-regulated by histamine.However,examination by confocal microscope revealed no detectable alterations in the morphology of the tight junctions after stimulating with histamine.CONCLUSION Our findings that histamine could regulate the mRNA expression of TJs of bronchial epithelial cells.That might contribute to the pathogenesis of chronic airway inflammatory diseases.