Division of Biomedical Sciences, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK.
Am J Physiol Cell Physiol. 2013 Apr 15;304(8):C739-47. doi: 10.1152/ajpcell.00164.2012. Epub 2013 Jan 30.
This study investigated the molecular identity and impact of enhancing voltage-gated Na(+) (Na(V)) channels in the control of vascular tone. In rat isolated mesenteric and femoral arteries mounted for isometric tension recording, the vascular actions of the Na(V) channel activator veratridine were examined. Na(V) channel expression was probed by molecular techniques and immunocytochemistry. In mesenteric arteries, veratridine induced potent contractions (pEC(50) = 5.19 ± 0.20, E(max) = 12.0 ± 2.7 mN), which were inhibited by 1 μM TTX (a blocker of all Na(V) channel isoforms, except Na(V)1.5, Na(V)1.8, and Na(V)1.9), but not by selective blockers of Na(V)1.7 (ProTx-II, 10 nM) or Na(V)1.8 (A-80347, 1 μM) channels. The responses were insensitive to endothelium removal but were partly (~60%) reduced by chemical destruction of sympathetic nerves by 6-hydroxydopamine (2 mM) or antagonism at the α1-adrenoceptor by prazosin (1 μM). KB-R7943, a blocker of the reverse mode of the Na(+)/Ca(2+) exchanger (3 μM), inhibited veratridine contractions in the absence or presence of prazosin. T16A(inh)-A01, a Ca(2+)-activated Cl(-) channel blocker (10 μM), also inhibited the prazosin-resistant contraction to veratridine. Na(V) channel immunoreactivity was detected in freshly isolated mesenteric myocytes, with apparent colocalization with the Na(+)/Ca(2+) exchanger. Veratridine induced similar contractile effects in the femoral artery, and mRNA transcripts for Na(V)1.2 and Na(V)1.3 channels were evident in both vessel types. We conclude that, in addition to sympathetic nerves, NaV channels are expressed in vascular myocytes, where they are functionally coupled to the reverse mode of Na(+)/Ca(2+) exchanger and subsequent activation of Ca(2+)-activated Cl(-) channels, causing contraction. The TTX-sensitive Na(V)1.2 and Na(V)1.3 channels are likely involved in vascular control.
本研究旨在探究增强电压门控钠离子通道(Na(V))对血管张力控制的分子机制及其影响。本研究采用大鼠离体肠系膜和股动脉进行等长张力记录,检测了 Na(V)通道激活剂藜芦碱的血管作用。通过分子技术和免疫细胞化学方法探测 Na(V)通道表达。在肠系膜动脉中,藜芦碱诱导产生强烈的收缩反应(pEC(50) = 5.19 ± 0.20,E(max) = 12.0 ± 2.7 mN),该反应可被 1 μM TTX(除 Na(V)1.5、Na(V)1.8 和 Na(V)1.9 以外所有 Na(V)通道同工型的阻滞剂)所抑制,但不能被 Na(V)1.7(ProTx-II,10 nM)或 Na(V)1.8(A-80347,1 μM)通道的选择性阻滞剂所抑制。该反应不受血管内皮去除的影响,但可被 6-羟多巴胺(2 mM)化学破坏交感神经或普萘洛尔(1 μM)拮抗α1-肾上腺素受体而部分(约 60%)抑制。Na(+)/Ca(2+)交换的反向模式阻滞剂 KB-R7943(3 μM)可抑制普萘洛尔存在或不存在时藜芦碱引起的收缩反应。Ca(2+)激活的 Cl(-)通道阻滞剂 T16A(inh)-A01(10 μM)也抑制了对普萘洛尔耐药的藜芦碱收缩反应。在新鲜分离的肠系膜肌细胞中检测到 Na(V)通道免疫反应性,与 Na(+)/Ca(2+)交换器有明显的共定位。藜芦碱在股动脉中也引起类似的收缩作用,两种血管类型均存在 Na(V)1.2 和 Na(V)1.3 通道的 mRNA 转录本。我们得出结论,除了交感神经外,Na(V)通道也在血管肌细胞中表达,在这些细胞中,它们与 Na(+)/Ca(2+)交换的反向模式和随后的 Ca(2+)激活的 Cl(-)通道激活功能耦联,引起收缩。TTX 敏感的 Na(V)1.2 和 Na(V)1.3 通道可能参与血管控制。