Dept. of Molecular Pharmacology & Therapeutics, Loyola Univ. Chicago, Stritch School of Medicine, 2160 S. First Ave., Bldg. 102, Rm. 3634, Maywood, IL 60153.
Am J Physiol Lung Cell Mol Physiol. 2014 Mar 15;306(6):L476-86. doi: 10.1152/ajplung.00253.2013. Epub 2014 Jan 17.
KCNQ (Kv7 family) potassium (K(+)) channels were recently found in airway smooth muscle cells (ASMCs) from rodent and human bronchioles. In the present study, we evaluated expression of KCNQ channels and their role in constriction/relaxation of rat airways. Real-time RT-PCR analysis revealed expression of KCNQ4 > KCNQ5 > KCNQ1 > KCNQ2 > KCNQ3, and patch-clamp electrophysiology detected KCNQ currents in rat ASMCs. In precision-cut lung slices, the KCNQ channel activator retigabine induced a concentration-dependent relaxation of small bronchioles preconstricted with methacholine (MeCh; EC50 = 3.6 ± 0.3 μM). Bronchoconstriction was also attenuated in the presence of two other structurally unrelated KCNQ channel activators: zinc pyrithione (ZnPyr; 1 μM; 22 ± 7%) and 2,5-dimethylcelecoxib (10 μM; 24 ± 8%). The same three KCNQ channel activators increased KCNQ currents in ASMCs by two- to threefold. The bronchorelaxant effects of retigabine and ZnPyr were prevented by inclusion of the KCNQ channel blocker XE991. A long-acting β2-adrenergic receptor agonist, formoterol (10 nM), did not increase KCNQ current amplitude in ASMCs, but formoterol (1-1,000 nM) did induce a time- and concentration-dependent relaxation of rat airways, with a notable desensitization during a 30-min treatment or with repetitive treatments. Coadministration of retigabine (10 μM) with formoterol produced a greater peak and sustained reduction of MeCh-induced bronchoconstriction and reduced the apparent desensitization observed with formoterol alone. Our findings support a role for KCNQ K(+) channels in the regulation of airway diameter. A combination of a β2-adrenergic receptor agonist with a KCNQ channel activator may improve bronchodilator therapy.
KCNQ(Kv7 家族)钾(K(+))通道最近在啮齿动物和人类小支气管的气道平滑肌细胞(ASMCs)中被发现。在本研究中,我们评估了 KCNQ 通道的表达及其在大鼠气道收缩/松弛中的作用。实时 RT-PCR 分析显示 KCNQ4>KCNQ5>KCNQ1>KCNQ2>KCNQ3 的表达,膜片钳电生理学检测到大鼠 ASMCs 中的 KCNQ 电流。在精密切割肺切片中,KCNQ 通道激活剂 retigabine 诱导预先用乙酰甲胆碱(MeCh;EC50=3.6±0.3μM)预收缩的小支气管浓度依赖性松弛。在存在另外两种结构上不相关的 KCNQ 通道激活剂的情况下,支气管收缩也被减弱:锌吡啶硫酮(ZnPyr;1μM;22±7%)和 2,5-二甲基塞来昔布(10μM;24±8%)。这三种 KCNQ 通道激活剂都将 ASMCs 中的 KCNQ 电流增加了两到三倍。Retigabine 和 ZnPyr 的支气管舒张作用被 KCNQ 通道阻滞剂 XE991 的加入所阻止。长效β2-肾上腺素能受体激动剂福莫特罗(10nM)不会增加 ASMCs 中的 KCNQ 电流幅度,但福莫特罗(1-1000nM)确实会引起大鼠气道的时间和浓度依赖性松弛,在 30 分钟的治疗或重复治疗期间观察到明显的脱敏。Retigabine(10μM)与福莫特罗同时给药可产生更大的峰值和持续降低乙酰甲胆碱诱导的支气管收缩,并减少单独使用福莫特罗时观察到的明显脱敏。我们的研究结果支持 KCNQ K(+)通道在气道直径调节中的作用。β2-肾上腺素能受体激动剂与 KCNQ 通道激活剂的联合应用可能改善支气管扩张剂治疗。