Department of Molecular Pharmacology & Therapeutics, Loyola University Chicago, Maywood, IL 60153, USA.
Am J Physiol Lung Cell Mol Physiol. 2012 Jan 1;302(1):L120-32. doi: 10.1152/ajplung.00194.2011. Epub 2011 Sep 30.
Expression and function of Kv7 (KCNQ) voltage-activated potassium channels in guinea pig and human airway smooth muscle cells (ASMCs) were investigated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), patch-clamp electrophysiology, and precision-cut lung slices. qRT-PCR revealed expression of multiple KCNQ genes in both guinea pig and human ASMCs. Currents with electrophysiological and pharmacological characteristics of Kv7 currents were measured in freshly isolated guinea pig and human ASMCs. In guinea pig ASMCs, Kv7 currents were significantly suppressed by application of the bronchoconstrictor agonists methacholine (100 nM) or histamine (30 μM), but current amplitudes were restored by addition of a Kv7 channel activator, flupirtine (10 μM). Kv7 currents in guinea pig ASMCs were also significantly enhanced by another Kv7.2-7.5 channel activator, retigabine, and by celecoxib and 2,5-dimethyl celecoxib. In precision-cut human lung slices, constriction of airways by histamine was significantly reduced in the presence of flupirtine. Kv7 currents in both guinea pig and human ASMCs were inhibited by the Kv7 channel blocker XE991. In human lung slices, XE991 induced robust airway constriction, which was completely reversed by addition of the calcium channel blocker verapamil. These findings suggest that Kv7 channels in ASMCs play an essential role in the regulation of airway diameter and may be targeted pharmacologically to relieve airway hyperconstriction induced by elevated concentrations of bronchoconstrictor agonists.
通过定量逆转录聚合酶链反应(qRT-PCR)、膜片钳电生理学和离体鼠肺切片研究了豚鼠和人气道平滑肌细胞(ASMCs)中 Kv7(KCNQ)电压激活钾通道的表达和功能。qRT-PCR 显示豚鼠和人 ASMCs 中多个 KCNQ 基因的表达。在新鲜分离的豚鼠和人 ASMCs 中测量具有 Kv7 电流电生理和药理学特征的电流。在豚鼠 ASMCs 中,Kv7 电流应用支气管收缩剂激动剂(如乙酰甲胆碱(100 nM)或组胺(30 μM)时显著受到抑制,但电流幅度通过添加 Kv7 通道激活剂(如氟比洛芬(10 μM))得到恢复。在豚鼠 ASMCs 中,另一种 Kv7.2-7.5 通道激活剂(瑞替加滨)以及塞来昔布和 2,5-二甲基塞来昔布也显著增强了 Kv7 电流。在离体人肺切片中,在氟比洛芬存在的情况下,组胺引起的气道收缩明显减少。Kv7 通道阻断剂 XE991 抑制豚鼠和人 ASMCs 中的 Kv7 电流。在人肺切片中,XE991 诱导强烈的气道收缩,加入钙通道阻断剂维拉帕米后完全逆转。这些发现表明,ASMCs 中的 Kv7 通道在气道直径的调节中起着重要作用,并且可能成为药理学靶点,以缓解由升高的支气管收缩剂浓度引起的气道过度收缩。