流感病毒对囊性纤维化跨膜转导调节因子影响的研究  

Influences of influenza virus on cystic fibrosis transmembrane conductance regulator （CFTR）

作　　者：候亚鹏 周祉妤 昌建钧 丁炎[1] 聂宏光[1] 

机构地区：[1]中国医科大学代谢病分子机制与药物研究所,沈阳110122

出　　处：《中华微生物学和免疫学杂志》2017年第11期805-809,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（81670010,81270098）;辽宁省高等学校基本科研项目（LQNK201745）

摘　　要：目的 探讨原代培养的小鼠气管单层细胞培养方法及流感病毒对原代培养的小鼠气管单层细胞囊性纤维化跨膜转导调节因子的影响，明确流感病毒破坏呼吸系统水盐转运平衡的影响机制。方法 应用倒置显微镜记录细胞形态，应用电阻计测量跨上皮电阻以评估细胞间的紧密连接状态，尤斯灌流装置记录原代培养的小鼠气管单层细胞短路电流。结果 200倍焦距的光镜下培养的原代小鼠气管上皮细胞形态呈紧密连接的铺路石状，跨上皮电阻在培养6 d后大于1 000 Ω；在细胞膜完整的条件下，流感病毒能降低囊性纤维化跨膜转导调节因子短路电流至（52.77±10.30）%，在基底膜通透的细胞单层中，流感病毒能降低囊性纤维化跨膜转导调节因子短路电流至（41.50±1.09）%，而囊性纤维跨膜转导调节因子在调节呼吸道水盐转运平衡中有重要作用。结论 气液交界面模式培养小鼠气管上皮细胞非常高效，并且接近小鼠正常的生理状态；流感病毒可能通过抑制囊性纤维化跨膜转导调节因子的功能，阻滞阴离子分泌，进而诱发慢性阻塞性肺病以及哮喘等肺部相关性疾病。Objective To evaluate the differentiation of mouse tracheal epithelial cells （MTEC） at an air-liquid interface and to investigate the influences of influenza virus on the cystic fibrosis transmem- brahe conductance regulator （CFFR） in primary cultured MTEC for further elucidating the possible mecha- nism of imbalanced fluid and salt transportation in respiratory system caused by influenza virus infection. Methods The morphology of primarily cultured MTEC was observed under inverted microscope. Trans epi- thelial electrical resistance （TEER） was measured by a resistance meter to evaluate the integrity of cultured MTEC. An Ussing chamber apparatus was used to record the short-circuit current of primary cultured MTEC. Results The primarily cultured MTEC clustered together and had a tight pavement-like appearance under light microscope. The TEER was greater than 1 000 fl after 6 days of culture. Influenza virus could reduce the short-circuit current of CFTR to （52.77±10.30）% in intact cell membrane and to （41.50±1.09）% in monolayer MTEC after increasing the permeability of basement membrane. It had been proved that CFTR was essential to maintaining the balance of fluid and salt transportation in respiratory system. Conclusion Mouse MTEC are efficiently cultured at a air-liquid interface and the primarily cultured cells are highly similar to those in a normal physiologic state. Influenza virus may block the secretion of anions through inhibiting the function of CFTR, which may induce the development of chronic obstructive pulmonary disease and the incidence of asthma.

关 键 词：流感病毒 小鼠气管上皮细胞 囊性纤维化跨膜转导调节因子:短路电流 

分 类 号：R373[医药卫生—病原生物学]