小鼠呼吸道原代上皮细胞分泌三磷酸腺苷的特征研究

Research on characteristics of adenosine triphosphate secreted by primary airway epithelial cells in mice

目的研究呼吸道上皮细胞分泌的三磷酸腺苷(ATP)与呼吸道清除功能密切相关,建立小动物呼吸道上皮原代细胞培养方法,比较分析其分泌ATP的特征。方法采用酶消化法制备小鼠支气管呼吸道上皮原代细胞,在低渗液刺激后的不同时间,用生化方法直接测定培养液上清中ATP含量。结果培养的呼吸道原代细胞可传代10代以上。原代细胞在低渗处理后,可快速产生ATP,其最大分泌量略低于正常人肺上皮细胞系,但产生速度较快。此方法建立的细胞对低渗刺激具有更真实的反应性,可用于上皮细胞的功能研究。结论原代呼吸道上皮细胞具有产生ATP的能力,其特征与上皮细胞系和原代平滑肌细胞产生ATP的特征有所不同。气道上皮和平滑肌细胞可同时产生ATP,有助于协同排除异物和病原体。

Objective To establish the culturing method of primary airway epithelial cells of little animals in order to study the relationship between adenosine triphosphate （ ATP ） and cleaning function of the respiratory tract, and comparatively analyze the characteristics of ATP secreted by primary airway epithelial cells. Methods Enzyme digestion method was used to prepare primary bronchial airway epithelial cells from mice. At different time after hypotonic solution stimulation, biochemical method was used to directly detect the ATP content in the supematant of the culture solution. Results The cultured primary epithelial cells were able to pass more than 10 generations. After the hypotonic treatment, the primary cells could rapidly secrete ATP. The maximum secretory content was only lower than that of healthy humang pulmonary epithelial cells, but the secretory speed of primary cells was faster. The cells prepared by this method had realer reactiveness to hypotonic stimulation, which could be used in the function research on epithelial cells. Conclusion Primary airway epithelial cells have the ability to secrete ATP. Its characteristics are different from the pulmonary epithelial cells and primary bronchial1 smooth muscle cells. The epithelia of airway and the smooth muscle cells can secrete ATP at the same time, which is helpful to eliminate pathogens.