Cystic fibrosis decreases the apical membrane chloride permeability of monolayers cultured from cells of tracheal epithelium.
Cystic fibrosis decreases the apical membrane chloride permeability of monolayers cultured from cells of tracheal epithelium.
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囊性纤维化降低了气管上皮细胞培养的单层细胞的顶膜氯离子渗透性。
DOI：
10.1073/pnas.82.18.6167
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发表时间：
1985
影响因子：
11.1
通讯作者：
Finkbeiner,WE
中科院分区：
文献类型：
--
作者：
Widdicombe,JH;Welsh,MJ;Finkbeiner,WE
一名12岁女孩死于囊性纤维化后4小时用胶原酶消化气管粘膜。获得了4000万个活细胞。将细胞以10（6）/cm 2接种到4个涂有人胎盘胶原蛋白的Nuclepore滤器上，1天后形成融合单层。其超微结构与正常人细胞相似。在接种后第5-7天，在常规的Ussing室中或用细胞内微电极对其进行研究。单层显示出380 +/-50 Ω X cm 2的电阻和1.8 +/-0.4 microA X cm-2的短路电流（Isc）。该抗性与犬或正常人单层细胞获得的抗性相似。Isc小于正常人（约3 microA X cm-2）或狗（约10 microA X cm-2）细胞。囊性纤维化细胞类似于正常的单层，因为沙丁胺醇哇巴因和粘膜阿米洛利抑制Isc，而粘膜哇巴因或沙丁胺醇阿米洛利没有影响。它们与正常人或狗细胞的不同之处在于，布美他尼不抑制Isc，异丙肾上腺素或前列腺素E2对Isc的刺激大大减少或消除。另外异丙肾上腺素去极化顶端膜电位（psi a）和降低分数电阻（fR）在正常人和狗，但没有影响的psi a或fR在囊性纤维化细胞。通过用葡萄糖酸盐替代将粘膜氯化物从120 mM降低至5 mM，使犬和正常人单层的fR增加，去极化psi a增加22（犬）或30（人）mV。在囊性纤维化单层，氯离子置换超极化psi一个2 mV，并对fR的影响不大。这些结果表明，在囊性纤维化的主要缺陷是减少顶端膜氯离子电导。
The tracheal mucosa from a 12-year-old girl was digested with collagenase 4 hr after her death from cystic fibrosis. Forty million viable cells were obtained. The cells, plated at 10(6) per cm2 onto four Nuclepore filters coated with human placental collagen, formed confluent monolayers after 1 day. Their ultrastructure was similar to that of normal human cells. They were studied in conventional Ussing chambers or with intracellular microelectrodes on days 5-7 after plating. The monolayers displayed resistance of 380 +/- 50 omega X cm2 and short-circuit current (Isc) of 1.8 +/- 0.4 microA X cm-2. This resistance is similar to that obtained for dog or normal human monolayers. The Isc is less than normal human (approximately 3 microA X cm-2) or dog (approximately 10 microA X cm-2) cells. The cystic fibrosis cells resembled normal monolayers in that serosal ouabain and mucosal amiloride inhibited Isc, while mucosal ouabain or serosal amiloride had no effect. They differed from normal human or dog cells in that Isc was not inhibited by bumetanide and the stimulation of Isc by isoproterenol or prostaglandin E2 was greatly reduced or abolished. Addition of isoproterenol depolarized apical membrane potential (psi a) and decreased fractional resistance (fR) in normal human and dog but had no effect on psi a or fR in cystic fibrosis cells. Reduction of mucosal chloride from 120 to 5 mM by replacement with gluconate increased fR of dog and normal human monolayers and depolarized psi a by 22 (dog) or 30 (human) mV. In cystic fibrosis monolayers, chloride replacement hyperpolarized psi a by 2 mV and had little effect on fR. These results suggest that the primary defect in cystic fibrosis is reduced apical membrane chloride conductance.