Activation of CFTR trafficking and gating by vasoactive intestinal peptide in human bronchial epithelial cells
Activation of CFTR trafficking and gating by vasoactive intestinal peptide in human bronchial epithelial cells
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DOI：
10.1002/jcb.22999
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发表时间：
2011-03
影响因子：
4
通讯作者：
F. Qu;Hui-jun Liu;Y. Xiang;Yu-Rong Tan;Chi Liu;Xiao-lin Zhu;X. Qin
中科院分区：
文献类型：
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作者：
F. Qu;Hui-jun Liu;Y. Xiang;Yu-Rong Tan;Chi Liu;Xiao-lin Zhu;X. Qin
囊性纤维化跨膜传导调节因子（CFTR）是一种顶端膜氯离子通道，对调节上皮细胞和其他细胞类型中的液体、氯离子和碳酸氢盐转运至关重要。囊性纤维化（CF）的最常见原因是CFTR突变体的异常运输。因此，理解将CFTR从内质网转运到细胞表面的细胞机制是重要的。血管活性肠多肽（VIP）在CFTR依赖性氯离子转运中起重要作用。本研究旨在观察VIP对人支气管上皮细胞CFTR转运及通道门控的影响。共聚焦显微镜显示CFTR免疫荧光从顶膜深入细胞质。VIP处理后，CFTR免疫荧光向细胞的顶端延伸减少，CFTR荧光的峰强度向顶端膜移动。Western blot显示VIP增加细胞表面和总CFTR。与总CFTR增加水平相比，表面CFTR增加更明显。免疫沉淀法发现VIP处理的HBEC中CFTR的成熟形式显著增加。VIP导致HBEC中Cl−流出量增加三倍。VIP（10−8 mol/L）刺激后，始终观察到格列本脲敏感性和DIDS不敏感性CFTR Cl−电流。VIP（10−8 mol/L）增强的CFTR Cl−电流的增加至少部分被蛋白激酶A（PKA）抑制剂H-89和蛋白激酶C（PKC）抑制剂H-7逆转，表明PKA和PKC参与VIP促进的CFTR Cl −电流。J.细胞。112：902-908，2011.© 2010 Wiley利斯公司
Cystic fibrosis transmembrane conductance regulator (CFTR) is an apical membrane chloride channel critical to the regulation of fluid, chloride, and bicarbonate transport in epithelia and other cell types. The most common cause of cystic fibrosis (CF) is the abnormal trafficking of CFTR mutants. Therefore, understanding the cellular machineries that transit CFTR from the endoplasmic reticulum to the cell surface is important. Vasoactive intestinal polypeptide (VIP) plays an important role in CFTR‐dependent chloride transport. The present study was designed to observe the affection of VIP on the trafficking of CFTR, and channel gating in human bronchial epithelium cells (HBEC). Confocal microscopy revealed CFTR immunofluorescence extending from the apical membrane deeply into the cell cytoplasm. After VIP treatment, apical extension of CFTR immunofluorescence into the cell was reduced and the peak intensity of CFTR fluorescence shifted towards the apical membrane. Western blot showed VIP increased cell surface and total CFTR. Compared with the augmented level of total CFTR, the surface CFTR increased more markedly. Immunoprecipitation founded that the mature form of CFTR had a marked increase in HBEC treated with VIP. VIP led to a threefold increase in Cl− efflux in HBEC. Glibenclamide‐sensitive and DIDS‐insensitive CFTR Cl− currents were consistently observed after stimulation with VIP (10−8 mol/L). The augmentation of CFTR Cl− currents enhanced by VIP (10−8 mol/L) was reversed, at least in part, by the protein kinase A (PKA) inhibitor, H‐89 and the protein kinase C (PKC) inhibitor, H‐7, suggesting PKA and PKC participate in the VIP‐promoted CFTR Cl− currents. J. Cell. Biochem. 112: 902–908, 2011. © 2010 Wiley‐Liss, Inc.