Ciliary beating amplitude controlled by intracellular Cl- and a high rate of CO2 production in ciliated human nasal epithelial cells
Ciliary beating amplitude controlled by intracellular Cl- and a high rate of CO2 production in ciliated human nasal epithelial cells
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纤毛人鼻上皮细胞中细胞内 Cl- 控制纤毛跳动幅度和高 CO2 产生率
DOI：
10.1007/s00424-019-02280-5
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Nakahari T
中科院分区：
文献类型：
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作者：
Inui TA;Murakami K;Yasuda M;Hirano S;Ikeuchi Y;Kogiso H;Hosogi S;Inui T;MARUNAKA Y;Nakahari T
纤毛运动受纤毛搏动频率和振幅两个参数控制。在这项研究中，我们开发了一种新的方法来测量CBF和纤毛弯曲距离（CBD，纤毛跳动幅度的指数）在纤毛人鼻上皮细胞（cHNECs）的原代培养，这是从患者的过敏性鼻炎和慢性鼻窦炎的合同准备。无Cl− NO3−溶液或bumetanide（Na+/K+/2Cl−共转运抑制剂）的应用，降低细胞内Cl−浓度（[Cl−]i），在37 °C时增加CBD，而不是CBF;然而，在25 °C时增加CBD和CBF。相反，加入Cl−通道阻滞剂（5-硝基-2-（3-苯基丙基氨基）苯甲酸（NPPB）和4-[[4-氧代-2-硫代-3-[3-三氟甲基]苯基]-5-噻唑烷亚基]甲基]苯甲酸（CFTR（inh）-172）），增加[Cl−]i，降低CBD和CBF，表明CFTR在维持这些细胞中的[Cl−] i中起着至关重要的作用。我们推测Cl−调节cHNECs中CBD和CBF的分子马达的活动。此外，应用无CO2/HCO 3 −溶液不会改变细胞内pH值（pHi），加入碳酸酐酶抑制剂（乙酰唑胺）可维持NH 4+脉冲诱导的pH值升高，而在没有乙酰唑胺的情况下，NH 4+脉冲可短暂升高pH值。这些结果表明，cHNEC产生大量的CO2，即使在无CO2/HCO 3 −的条件下也能保持恒定的pH。
The ciliary transport is controlled by two parameters of the ciliary beating, frequency (CBF) and amplitude. In this study, we developed a novel method to measure both CBF and ciliary bend distance (CBD, an index of ciliary beating amplitude) in ciliated human nasal epithelial cells (cHNECs) in primary culture, which are prepared from patients contracting allergic rhinitis and chronic sinusitis. An application of Cl−-free NO3−solution or bumetanide (an inhibitor of Na+/K+/2Cl−cotransport), which decreases intracellular Cl−concentration ([Cl−]i), increased CBD, not CBF, at 37 °C; however, it increased both CBD and CBF at 25 °C. Conversely, addition of Cl−channel blockers (5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and 4-[[4-Oxo-2-thioxo-3-[3-trifluoromethyl]phenyl]-5-thiazolidinylidene]methyl] benzoic acid (CFTR(inh)-172)), which increase [Cl−]i, decreased both CBD and CBF, suggesting that CFTR plays a crucial role for maintaining [Cl−]iin these cells. We speculate that Cl−modulates activities of the molecular motors regulating both CBD and CBF in cHNECs. Moreover, application of the CO2/HCO3−-free solution did not change intracellular pH (pHi), and addition of an inhibitor of carbonic anhydrase (acetazolamide) sustained pHiincrease induced by the NH4+pulse, which transiently increased pHiin the absence of acetazolamide. These results indicate that the cHNEC produces a large amount of CO2, which maintains a constant pHieven under the CO2/HCO3−-free condition.