EVIDENCE FOR REDUCED CL- AND INCREASED NA+ PERMEABILITY IN CYSTIC-FIBROSIS HUMAN PRIMARY-CELL CULTURES
EVIDENCE FOR REDUCED CL- AND INCREASED NA+ PERMEABILITY IN CYSTIC-FIBROSIS HUMAN PRIMARY-CELL CULTURES
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DOI：
10.1113/jphysiol.1988.sp017322
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发表时间：
1988-11-01
影响因子：
5.5
通讯作者：
YANKASKAS, JR
中科院分区：
文献类型：
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作者：
BOUCHER, RC;COTTON, CU;YANKASKAS, JR
1.采用原代细胞培养系统和细胞内微电极，对正常人和囊性纤维化(CF)人气道上皮细胞顶膜Na~+和Cl~-通路进行了定量检测和比较。2.与原位跨上皮差(PD)一样，培养的CF上皮的PD为(-27.+-)。4 mV，平均+-。S.E.M.(n=28)超过正常上皮的PD(-10.+-)。1 mV；n=22)。PD升高主要反映了CF上皮主动转运速率(相当于短路，IEQ)的增加(61.6+-)。9µA·cm~(-2)，与正常上皮(23±-)比较。3µA厘米-2)。在跨上皮阻力方面没有检测到显著差异。3.离子置换研究表明，正常细胞顶膜具有可被异丙肾上腺素激活的顶膜氯电导(GCL)。CF细胞不显示顶膜GCL，GCL也不能被异丙肾上腺素激活。4.与正常细胞相比，CF细胞对阿米洛利敏感的IEQ和对阿米洛利敏感的根尖膜电导(GNA)更大。此外，异丙肾上腺素可增加CF对阿米洛利的敏感性IEQ，但不能增加正常的呼吸道上皮细胞。5.等效电路分析表明，与正常细胞相比，CF细胞的顶膜电动势(EMF)较正，基底膜电势(EMF)较负。对于正常细胞和CF细胞，根尖(Ra)和基底侧(Rb)膜的基线阻力没有差别。6.通过等效电路分析或离子替代对细胞旁通路对离子流(Rs)的抵抗力的估计，发现CF和正常细胞的Rs没有差异。7.细胞内氯离子通透性降低和Na+通透性升高是培养的CF型呼吸道上皮细胞的特征。这些数据表明，顶膜通透性调节的缺陷是本病的主要特征。
1. Employing a primary cell culture system and intracellular microelectrodes, we quantitated and compared the Na+ and Cl- pathways in apical membranes of normal and cystic fibrosis (CF) human airway epithelia. 2. Like the transepithelial difference (PD) in situ, the PD of CF epithelia in culture (-27 .+-. 4 mV, mean .+-. S.E.M.; n = 28) exceeded the PD of normal epithelia (-10 .+-. 1 mV; n = 22). The raised PD principally reflected an increase in the rate of active transport (equivalent short circuit, Ieq) for CF epithelia (61 .+-. 9 .mu.A cm-2) as compared with normal epithelia (23 .+-. 3 .mu.A cm-2). No significant differences in transepithelial resistance were detected. 3. As indicated by ion replacement studies (gluconate for Cl-), the apical membrane of normal cells exhibits an apical membrane Cl- conductance (GCl) that can be activated by isoprenaline. CF cells do not exhibit an apical membrane GCl, nor can a GCl be activated by isoprenaline. 4. CF cells exhibited a larger amiloride-sensitive Ieq and amiloride-sensitive apical membrane conductance (GNa) than normal cells. Further, the amiloride-sensitive Ieq was increased by isoprenaline in CF but not normal airway epithelia. 5. Equivalent circuit analysis yielded evidence for a more positive electromotive force (EMF) across the apical membrane and a more negative EMF across the basolateral membrane of CF cells as compared with normal cells. Baseline resistances of the apical (Ra) and basolateral (Rb) membranes did not differ for normal and CF cells. 6. Estimates of the resistance of the paracellular path to ion flow (Rs) by equivalent circuit analysis or ion substitution detected no differences in Rs between CF and normal cells. 7. We conclude that abnormalities in both cellular Cl-1 permeability (reduced) and Na+ permeability (increased) are characteristic of the cultured CF respiratory epithelial cell. These data suggest that a defect in the regulation of apical membrane permeabilities is a central feature of this disease.