CAJ | 학술논문

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mRNA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mRNA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mRNA表达，有可能是慢性呼吸道炎性疾病的发病机制之一。
목적검측조알자격하체외배양적원대지기관상피세포간긴밀련접상관기인표체적개변。방법응용기-액계면배양방법체외배양정상인화효천환자원대지기관상피세포，응용조알자격상피세포96 h，분별응용실시형광정량PCR화면역형광염색공취초현미경검측긴밀련접상관기인재mRNA급단백수평적표체。결과실시형광정량PCR결과현시조알자격가이유도모사긴밀련접상관기인mRNA표체적상조혹하조；면역형광염색공취초현미경검사미발현조알자격가이유도모사긴밀련접상관단백발생형태학적개변。결론조알가이조절인원대지기관상피세포긴밀련접상관기인mRNA표체，유가능시만성호흡도염성질병적발병궤제지일。
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.