Expression of acid-sensing ion channels in intestinal epithelial cells and their role in the regulation of duodenal mucosal bicarbonate secretion.
Expression of acid-sensing ion channels in intestinal epithelial cells and their role in the regulation of duodenal mucosal bicarbonate secretion.
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DOI：
10.1111/j.1748-1716.2010.02207.x
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发表时间：
2011-01
期刊：
影响因子：
--
通讯作者：
Dong H
中科院分区：
文献类型：
--
作者：
Dong X;Ko KH;Chow J;Tuo B;Barrett KE;Dong H
尽管酸敏感离子通道（ASICs）已在多种哺乳动物细胞中得到研究，但目前对于它们在肠上皮细胞中的存在情况知之甚少。因此，本研究的目的是探究ASIC异构体在肠上皮细胞中的表达和功能，特别是它们在酸刺激的十二指肠黏膜碳酸氢盐分泌（DMBS）中的生理作用。采用逆转录聚合酶链反应（RT - PCR）和数字式钙离子成像技术来确定ASICs在人肠上皮细胞系HT29细胞中的表达和功能。在体内对C57黑鼠测量酸刺激的DMBS，以研究ASICs在这一生理过程中的作用。在从小鼠和HT29细胞剥离的十二指肠黏膜中检测到ASIC1a的信使核糖核酸（mRNA）表达，同时还测量了这些细胞对细胞外酸中毒的胞质游离钙离子（[Ca²⁺]cyt）反应。在含Ca²⁺的溶液中，酸中毒以pH依赖的方式升高[Ca²⁺]cyt，在大约pH5.9时达到半数最大效应（pH0.5）。酸中毒诱导的[Ca²⁺]cyt升高被阿米洛利（一种ASIC阻滞剂）、SK&F96365（一种非选择性阳离子通道阻滞剂）或无Ca²⁺溶液显著抑制；但受U73122或硝苯地平的影响较小。在酸中毒升高[Ca²⁺]cyt之后，用三磷酸腺苷（ATP）刺激嘌呤能受体进一步增加了[Ca²⁺]cyt。此外，酸刺激的小鼠DMBS被阿米洛利显著减弱。因此，ASIC1a在肠上皮细胞中功能性表达，并且可能在这些细胞的[Ca²⁺]cyt稳态调节以及酸刺激的DMBS中发挥重要作用。
Although acid-sensing ion channels (ASICs) have been studied in a variety of mammalian cells, little is currently known about their presence in intestinal epithelial cells. Therefore, the aims of the present study were to investigate the expression and function of ASIC isoforms in intestinal epithelial cells, particularly their physiological role in the acid-stimulated duodenal mucosal bicarbonate secretion (DMBS). RT-PCR and digital Ca2+ imaging were used to determine the expression and function of ASICs in HT29 cells, a human intestinal epithelial cell line. The acid-stimulated DMBS was measured in C57 black mice in vivo to study the role of ASICs in this physiological process. mRNA expression of ASIC1a was detected in the duodenal mucosa stripped from mice and HT29 cells, in which cytoplasmic free Ca2+ ([Ca2+]cyt) in response to extracellular acidosis was also measured. In Ca2+-containing solutions, acidosis raised [Ca2+]cyt in a pH-dependent manner with a half-maximal effect (pH0.5) at approximately 5.9. The acidosis-induced increase in [Ca2+]cyt was markedly inhibited by amiloride (an ASIC blocker), SK&F96365 (a blocker for non-selective cation channels), or in Ca2+-free solutions; but slightly affected by U73122, or nifedipine. After acidosis raised [Ca2+]cyt, stimulation of purinergic receptors with ATP further increased [Ca2+]cyt. Moreover, the acid-stimulated murine DMBS was significantly attenuated by amiloride. Therefore, ASIC1a is functionally expressed in intestinal epithelial cells, and may play an important role in the regulation of [Ca2+]cyt homeostasis in these cells and the acid-stimulated DMBS.
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影响因子：
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作者：
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通讯作者：
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影响因子：
4.8
作者：
Lingueglia, E;deWeille, JR;Lazdunski, M
通讯作者：
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DOI：
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发表时间：
2007-12-21
影响因子：
3.1
作者：
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