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首页> 外文期刊>Cellular Physiology and Biochemistry >X-ray microanalysis of apical fluid in cystic fibrosis airway epithelial cell lines
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X-ray microanalysis of apical fluid in cystic fibrosis airway epithelial cell lines

机译：囊性纤维化气道上皮细胞系根尖液的X射线显微分析

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The ionic composition of the fluid lining the airways (airway surface liquid, ASL) in healthy subjects and patients with cystic fibrosis (CF) has been a matter of controversy. It has been attempted to resolve conflicting theories by using cell cultures, but published results show a wide variety of values for the ionic concentrations in the apical fluid in these cultures. To investigate CFTR-mediated HCO3 conductance and the role of HCO3-in regulating ASL pH we determined the pH of the fluid covering the apical surface of airway epithelial cells. A normal (16HBE14o-) and a CF (CFBE41o-) bronchial epithelial cell line were grown on membrane inserts in both a liquid-liquid interface culture system for 7 days, and in an air-liquid interface culture system for one month. The elemental composition of the fluid covering the apical surface was determined by X-ray microanalysis of frozen-hydrated specimens, or by X-ray microanalysis of Sephadex beads that had been equilibrated with the apical fluid. Analysis showed that the apical fluid had a Na+ and Cl+ concentration of about 80-100 mM and thus was slightly hypotonic. The ionic concentrations were somewhat higher in air-liquid interface than in liquid-liquid interface cultures. The apical fluid in CF cells had significantly higher concentrations of Na and Cl than that in control cultures. In control cultures, the concentrations of Na+ and Cl+ in the apical fluid increased if glibenclamide, an inhibitor of the cystic fibrosis transmembrane conductance regulator (CFTR) was added to the apical medium. Exposing the cells to the metabolic inhibitor NaCN also resulted in a significant increase of the Na+ and Cl+ concentrations in the apical fluid. The results agree with the notion that these cell cultures are mainly absorptive cells, and that ion absorption by the CF cells is reduced compared to that in normal cells. The pH measurements of the fluid covering the apical part of cell cultures support the notion that bicarbonate ions may be transported by CFTR, and that this can be inhibited by specific CFTR inhibitors. Copyright (c) 2006 S. Karger AG, Basel.
机译：在健康受试者和患有囊性纤维化（CF）的患者中，气道内的流体（气道表面液体，ASL）的离子组成一直是一个有争议的问题。已经尝试通过使用细胞培养物来解决矛盾的理论，但是公开的结果显示了这些培养物中顶液中离子浓度的各种值。为了研究CFTR介导的HCO3电导和HCO3-在调节ASL pH中的作用，我们确定了覆盖气道上皮细胞顶表面的液体的pH。正常（16HBE140-）和CF（CFBE410-）支气管上皮细胞系在液-液界面培养系统中的膜插入物上生长7天，并在气液界面培养系统中生长1个月。通过冷冻水合标本的X射线显微分析，或通过已经用顶液平衡的Sephadex珠的X射线显微分析，可以确定覆盖根尖表面的液体的元素组成。分析表明，根尖液的Na +和Cl +浓度约为80-100 mM，因此具有低渗性。气液界面中的离子浓度比液液界面中的离子浓度高。 CF细胞的顶液中Na和Cl的浓度比对照培养物中的高得多。在对照培养物中，如果将格列本脲（一种囊性纤维化跨膜电导调节剂（CFTR）的抑制剂）添加到顶端培养基中，则会增加顶端液中Na +和Cl +的浓度。将细胞暴露于代谢抑制剂NaCN也会导致根尖液中Na +和Cl +浓度显着增加。结果与以下观点一致：这些细胞培养物主要是吸收性细胞，与正常细胞相比，CF细胞的离子吸收减少。覆盖细胞培养物顶端的液体的pH测量值支持这样的观念，即碳酸氢根离子可以通过CFTR转运，并且可以被特定的CFTR抑制剂抑制。版权所有（c）2006 S.Karger AG，巴塞尔。

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