Dual Pathway Activated by tert-Butyl Hydroperoxide in Human Airway Anion Secretion
Dual Pathway Activated by tert-Butyl Hydroperoxide in Human Airway Anion Secretion
复制标题
DOI：
10.1124/jpet.108.141580
复制
发表时间：
2008-11
影响因子：
3.5
通讯作者：
T. Matsuno;Y. Ito;Takamasa Ohashi;M. Morise;N. Takeda;K. Shimokata;K. Imaizumi;H. Kume;Y. Hasegawa
中科院分区：
文献类型：
--
作者：
T. Matsuno;Y. Ito;Takamasa Ohashi;M. Morise;N. Takeda;K. Shimokata;K. Imaizumi;H. Kume;Y. Hasegawa
我们分析了由叔丁基过氧化氢（t-BOOH）诱导的离子转运的机制，叔丁基过氧化氢（t-BOOH）是一种膜渗透性氧化剂，已被广泛用作氧化应激的模型，在人气道上皮细胞（Calu-3）。我们发现，t-BOOH诱导的短路电流是由两个不同的组件，峰值组件（PC）和持续的组件（SC）。这两种组分都被H-89还原（N-[2-（4-溴肉桂氨基）乙基]-5-异喹啉）[10 μM，一种蛋白激酶A（PKA）抑制剂]和氯非铵（100 μM，一种cAMP依赖性K+通道抑制剂），但不受Charybdotoxin影响（50 nM，人中间电导Ca 2+激活的K+通道抑制剂），这表明PC和SC都是通过共同的PKA依赖性/Ca 2+非依赖性途径产生的。尽管如此，生理特性的分析表明，PC和SC归因于不同的途径。PC（而非SC）与顶端膜Cl-电导相关，并受到环氧合酶（考克斯）-2抑制剂NS-398（N-[2-（环己氧基）-4-硝基苯基]-甲磺酰胺; 10 μM）的抑制。相比之下，SC（而非PC）由对基底外侧Na+-K+-2Cl-协同转运蛋白（NKCC 1）抑制剂布美他尼（50 μM）敏感的组分组成，并被细胞松弛素D（10 μM）和（R）-（+）-反式-N-（4-吡啶基）-4-（1-氨乙基）-环己烷甲酰胺（Y-27632; 20 μM）诱导的细胞骨架功能障碍消除。总的来说，t-BOOH通过两个独立的途径诱导PKA相关阴离子分泌：通过考克斯-2依赖性/细胞色素依赖性途径快速激活顶端阴离子流出，通过考克斯-2依赖性/细胞色素依赖性途径相对延迟激活NKCC 1以进行基底外侧阴离子摄取。
We analyzed the mechanisms underlying the ion transport induced by tert-butyl hydroperoxide (t-BOOH), a membrane-permeant oxidant that has been widely used as a model of oxidative stress, in human airway epithelial cells (Calu-3). We found that t-BOOH induced a short-circuit current that was composed of two distinct components, a peaked component (PC) and a sustained component (SC). Both components were reduced by the presence of H-89 (N-[2-(4-bromocinnamylamino)ethyl]-5-isoquinoline) [10 μM, a protein kinase A (PKA) inhibitor] and clofilium (100 μM, a cAMP-dependent K+ channel inhibitor) but not by charybdotoxin (50 nM, a human intermediate conductance Ca2+-activated K+ channel inhibitor), suggesting that both PC and SC were generated through a common PKA-dependent/Ca2+-independent pathway. Notwithstanding, analyses of the physiological properties revealed that PC and SC were attributable to different pathways. PC, but not SC, was correlated with apical membrane Cl- conductance and was inhibited by the cyclooxygenase (COX)-2 inhibitor NS-398 (N-[2-(cyclohexyloxyl)-4-nitrophenyl]-methane sulfonamide; 10 μM). In contrast, SC, but not PC, was composed of a component sensitive to bumetanide (50 μM), an inhibitor of the basolateral Na+-K+-2Cl- cotransporter (NKCC1), and was abolished by the cytoskeleton dysfunction induced by cytochalasin D (10 μM) and (R)-(+)-trans-N-(4-pyridyl)-4-(1-aminoethyl)-cyclohexane carboxamide (Y-27632; 20 μM). Collectively, t-BOOH induces PKA-related anion secretion through two independent pathways: rapid activation of apical anion efflux through a COX-2-dependent/cytoskeleton-independent pathway and relatively delayed activation of NKCC1 for basolateral anion uptake through a COX-2-independent/cytoskeleton-dependent pathway.