Capsaicinoids Regulate Airway Anion Transporters through Rho Kinase- and Cyclic AMP-Dependent Mechanisms
Capsaicinoids Regulate Airway Anion Transporters through Rho Kinase- and Cyclic AMP-Dependent Mechanisms
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DOI：
10.1165/rcmb.2010-0332oc
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发表时间：
2011-10-01
影响因子：
6.4
通讯作者：
Hasegawa, Yoshinori
中科院分区：
文献类型：
--
作者：
Hibino, Yoshitaka;Morise, Masahiro;Hasegawa, Yoshinori
为了研究辣椒素对气道阴离子转运体的影响，我们记录并分析了人气道上皮细胞Calu-3的跨上皮电流。辣椒素（100 μ M）的应用衰减矢量阴离子转运，估计为短路电流（I-SC），前和后刺激毛喉素（10 μ M）与伴随的细胞溶质环磷酸腺苷（cAMP）水平的降低。辣椒素诱导的抑制I-SC也观察到8-溴-cAMP（1 mM，细胞可渗透的cAMP类似物）和3-异丁基-1-甲基黄嘌呤（1 mM，磷酸二酯酶的抑制剂）的反应。辣椒素诱导的I-SC抑制归因于抑制bumetanide（基底外侧Na+-K+-2 Cl-共转运体1的抑制剂）-和4，4 '-二硝基芪-2，2'-二磺酸（基底外侧HCO 3-依赖性阴离子转运体的抑制剂）敏感组分，其反映了通过基底外侧cAMP依赖性阴离子转运体的阴离子摄取。相反，辣椒素增强顶端Cl-传导，这反映了通过囊性纤维化跨膜传导调节器（cAMP调节的Cl-通道）的传导性。辣椒碱的所有这些自相矛盾的作用都被辣椒平模仿。毛喉素的应用也增加了磷酸化肌球蛋白磷酸酶靶亚基1，并且辣椒素和辣椒平阻止了磷酸化，这表明这些辣椒素类化合物承担Rho激酶抑制剂的方面。我们还发现，顶端Cl-电导的增量是由传统的Rho激酶抑制剂Y-27632（20 μ M）和HA-1077（20 μ M）引起的，具有对基底外侧Na+-K+-2 Cl-协同转运蛋白1的选择性抑制。总的来说，辣椒素通过下调基底外侧阴离子摄取来抑制cAMP介导的阴离子转运，矛盾的是伴随着顶端囊性纤维化跨膜电导调节剂介导的阴离子电导的上调。后者是通过抑制Rho激酶介导的，Rho激酶被认为与肌动蛋白细胞骨架相互作用。
To investigate the effects of capsaicinoids on airway anion transporters, we recorded and analyzed transepithelial currents inhuman airway epithelial Calu-3 cells. Application of capsaicin (100 mu M) attenuated vectorial anion transport, estimated as short-circuit currents (I-SC), before and after stimulation by forskolin (10 mu M) with concomitant reduction of cytosolic cyclic AMP (cAMP) levels. The capsaicin-induced inhibition of I-SC was also observed in the response to 8-bromo-cAMP (1 mM, a cell-permeable cAMP analog) and 3-isobutyl-1-methylxanthine (1 mM, an inhibitor of phosphodiesterases). The capsaicin-induced inhibition of I-SC was attributed to suppression of bumetanide (an inhibitor of the basolateral Na+-K+-2 Cl- cotransporter 1)- and 4,4'-dinitrostilbene-2,2'-disulfonic acid (an inhibitor of basolateral HCO3--dependent anion transporters)sensitive components, which reflect anion uptake via basolateral cAMP-dependent anion transporters. In contrast, capsaicin potentiated apical Cl- conductance, which reflects conductivity through the cystic fibrosis transmembrane conductance regulator, a cAMP-regulated Cl- channel. All these paradoxical effects of capsaicin were mimicked by capsazepine. Forskolin application also increased phosphorylated myosin phosphatase target subunit 1, and the phosphorylation was prevented by capsaicin and capsazepine, suggesting that these capsaicinoids assume aspects of Rho kinase inhibitors. We also found that the increments in apical Cl- conductance were caused by conventional Rho kinase inhibitors, Y-27632 (20 mu M) and HA-1077 (20 mu M), with selective inhibition of basolateral Na+-K+-2 Cl- cotransporter 1. Collectively, capsaicinoids inhibit cAMP-mediated anion transport through down-regulation of basolateral anion uptake, paradoxically accompanied by up-regulation of apical cystic fibrosis transmembrane conductance regulator-mediated anion conductance. The latter is mediated by inhibition of Rho-kinase, which is believed to interact with actin cytoskeleton.