Tobias Orth, Anastasia Pyanova, Simon Lux, Peter Kaiser, Isabel Reinheimer, Daniel Løgstrup Nielsen, Josef Ali Khalid, Salomé Rognant, Thomas A. Jepps, Vladimir V. Matchkov, Rudolf Schubert
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Rat mesenteric arteries were studied using isometric myography, FURA-2 fluorimetry and proximity ligation assay. The BK channel blocker iberiotoxin potentiated methoxamine-induced contractions. The cardiotonic steroid, ouabain (10<sup>−5</sup> M), induced a contractile effect of IBTX at basal tension prior to methoxamine administration and enhanced the pro-contractile effect of IBTX on methoxamine-induced contractions. These facilitating effects of ouabain were prevented by the inhibition of either NCX or Src kinase. Furthermore, inhibition of NCX or Src kinase reduced the BK channel-mediated negative feedback regulation of arterial contraction. The effects of NCX and Src kinase inhibition were independent of each other. Co-localization of the Na/K-ATPase and the BK channel was evident. Our data suggest that BK channels limit ouabain-induced vasocontraction by a dual mechanism involving the NCX and Src kinase signaling. The data propose that the NCX and the Src kinase pathways, mediating the ouabain-induced activation of the BK channel, act in an independent manner.</p>","PeriodicalId":50455,"journal":{"name":"FASEB Journal","volume":null,"pages":null},"PeriodicalIF":4.4000,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202400628RR","citationCount":"0","resultStr":"{\"title\":\"Vascular smooth muscle BK channels limit ouabain-induced vasocontraction: Dual role of the Na/K-ATPase as a hub for Src-kinase and the Na/Ca-exchanger\",\"authors\":\"Tobias Orth, Anastasia Pyanova, Simon Lux, Peter Kaiser, Isabel Reinheimer, Daniel Løgstrup Nielsen, Josef Ali Khalid, Salomé Rognant, Thomas A. Jepps, Vladimir V. 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引用次数: 0
摘要
血管平滑肌中普遍存在大传导钙激活钾通道(BK 通道)和 Na/K-ATP 酶。Na/K-ATPase 可通过 Na/Ca 交换器(NCX)和 Src 激酶介导的细胞内 Ca2+ 浓度变化发挥作用。已知这两种途径都能调节 BK 通道。BK 通道在血管平滑肌细胞中是否与 Na/K-ATP 酶发生功能性相互作用仍有待阐明。因此,本研究探讨了 BK 通道限制乌苯那敏诱导的血管收缩的假设。研究人员使用等速肌电图、FURA-2荧光测定法和近距离结扎法对大鼠肠系膜动脉进行了研究。BK 通道阻断剂依比妥毒素增强了甲氧胺诱导的收缩。在给予甲氧胺之前,强心类固醇乌巴因(10-5 M)可在基础张力下诱导 IBTX 的收缩效应,并增强 IBTX 对甲氧胺诱导收缩的促进收缩效应。抑制 NCX 或 Src 激酶可阻止乌巴因的这些促进作用。此外,抑制 NCX 或 Src 激酶可减少 BK 通道介导的动脉收缩负反馈调节。抑制NCX和Src激酶的作用是相互独立的。Na/K-ATP 酶和 BK 通道的共定位是显而易见的。我们的数据表明,BK 通道通过 NCX 和 Src 激酶信号传导的双重机制限制了乌苯那敏诱导的血管收缩。这些数据表明,介导乌巴因诱导的 BK 通道激活的 NCX 和 Src 激酶途径以独立的方式发挥作用。
Vascular smooth muscle BK channels limit ouabain-induced vasocontraction: Dual role of the Na/K-ATPase as a hub for Src-kinase and the Na/Ca-exchanger
Large-conductance, calcium-activated potassium channels (BK channels) and the Na/K-ATPase are expressed universally in vascular smooth muscle. The Na/K-ATPase may act via changes in the intracellular Ca2+ concentration mediated by the Na/Ca exchanger (NCX) and via Src kinase. Both pathways are known to regulate BK channels. Whether BK channels functionally interact in vascular smooth muscle cells with the Na/K-ATPase remains to be elucidated. Thus, this study addressed the hypothesis that BK channels limit ouabain-induced vasocontraction. Rat mesenteric arteries were studied using isometric myography, FURA-2 fluorimetry and proximity ligation assay. The BK channel blocker iberiotoxin potentiated methoxamine-induced contractions. The cardiotonic steroid, ouabain (10−5 M), induced a contractile effect of IBTX at basal tension prior to methoxamine administration and enhanced the pro-contractile effect of IBTX on methoxamine-induced contractions. These facilitating effects of ouabain were prevented by the inhibition of either NCX or Src kinase. Furthermore, inhibition of NCX or Src kinase reduced the BK channel-mediated negative feedback regulation of arterial contraction. The effects of NCX and Src kinase inhibition were independent of each other. Co-localization of the Na/K-ATPase and the BK channel was evident. Our data suggest that BK channels limit ouabain-induced vasocontraction by a dual mechanism involving the NCX and Src kinase signaling. The data propose that the NCX and the Src kinase pathways, mediating the ouabain-induced activation of the BK channel, act in an independent manner.
期刊介绍:
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