Cuthbert A W, MacVinish L J
Department of Medicine, University of Cambridge, Addenbrooke's Hospital (Level 5, Box 157), Hills Road, Cambridge CB2 2QQ.
Br J Pharmacol. 2003 Sep;140(1):81-90. doi: 10.1038/sj.bjp.0705403. Epub 2003 Jul 14.
(1) Cultured epithelial monolayers of Calu-3 human airway cells were used to measure anion secretion in response to a number of phenanthrolines and benzoquinolines, using short-circuit current measurements. Calu-3 cells are derived from serous cells of submucosal glands of the airways and are a target for conditions in which muco-ciliary clearance is compromised. (2) Compounds studied were 5,6-benzoquinoline, 5-chloro-1,10-phenanthroline, 7,8-benzoquinoline, 5-nitro-1,10-phenanthroline, benzo[c]cinnoline and 1,10-phenanthroline, which gave EC50 values of 34, 48, 123, 235, 192 and 217 microm, respectively. Of these, 7,8-benzoquinoline was chosen for further detailed study. Concentration-response relationships for all the compounds had Hill slopes greater than 1. (3) Permeabilisation of the apical surface of epithelia with nystatin in the presence of an apical to basolateral potassium ion gradient reduced the EC50 for 7,8-benzoquinoline to 31 microm and altered the Hill slope to close to 1. (4) Using apically permeabilised epithelia it was shown that 7,8-benzoquinoline activates an intermediate-conductance calcium-sensitive potassium channel (KCNN4) and a cAMP-sensitive potassium channel (KCNQ1/KCNE3) in the basolateral epithelial membranes. (5) 7,8-Benzoquinoline was shown to increase chloride conductance of apical epithelial membranes, presumed to be by activation of the cystic fibrosis transmembrane conductance regulator. (6) 7,8-Benzoquinoline had a minor effect on cAMP accumulation in Calu-3 cells, probably by inhibition of phosphodiesterase, which may contribute to its effect on CFTR- and cAMP-sensitive potassium channels. (7) The usefulness of these novel actions in promoting secretion in airway submucosal glands is discussed.
(1) 使用短路电流测量法,以培养的Calu-3人呼吸道细胞单层来测量对多种菲咯啉和苯并喹啉的阴离子分泌反应。Calu-3细胞源自气道黏膜下腺的浆液细胞,是黏液纤毛清除功能受损病症的作用靶点。(2) 所研究的化合物有5,6-苯并喹啉、5-氯-1,10-菲咯啉、7,8-苯并喹啉、5-硝基-1,10-菲咯啉、苯并[c]噌啉和1,10-菲咯啉,其半数有效浓度(EC50)值分别为34、48、123、235、192和217微摩尔。其中,7,8-苯并喹啉被选作进一步详细研究对象。所有化合物的浓度-反应关系的希尔斜率均大于1。(3) 在存在从顶端到基底外侧的钾离子梯度的情况下,用制霉菌素通透上皮细胞的顶端表面,可使7,8-苯并喹啉的EC50降至31微摩尔,并使希尔斜率改变至接近1。(4) 利用顶端通透的上皮细胞表明,7,8-苯并喹啉可激活基底外侧上皮细胞膜中的一种中电导钙敏感钾通道(KCNN4)和一种cAMP敏感钾通道(KCNQ1/KCNE3)。(5) 7,8-苯并喹啉可增加顶端上皮细胞膜的氯电导,推测是通过激活囊性纤维化跨膜电导调节因子实现的。(6) 7,8-苯并喹啉对Calu-3细胞中cAMP积累有轻微影响,可能是通过抑制磷酸二酯酶,这可能有助于其对CFTR和cAMP敏感钾通道的作用。(7) 讨论了这些新作用在促进气道黏膜下腺分泌方面的实用性。