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首页> 外文期刊>Cellular Physiology and Biochemistry >Vectorial bicarbonate transport by capan-1 cells: A model for human pancreatic ductal secretion
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Vectorial bicarbonate transport by capan-1 cells: A model for human pancreatic ductal secretion

机译：capan-1细胞进行矢量碳酸氢盐转运：人胰管分泌模型

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摘要

Human pancreatic ducts secrete a bicarbonate-rich fluid but our knowledge of the secretory process is based mainly on studies of animal models. Our aim was to determine whether the HCO3- transport mechanisms in a human ductal cell line are similar to those previously identified in guinea-pig pancreatic ducts. Intracellular pH was measured by microfluorometry in Capan-1 cell monolayers grown on permeable filters and loaded with BCECF. Epithelial polarization was assessed by immunolocalization of occludin. Expression of mRNA for key electrolyte transporters and receptors was evaluated by RT-PCR. Capan-1 cells grown on permeable supports formed confluent, polarized monolayers with well developed tight junctions. The recovery of pH(i) from an acid load, induced by a short NH4+ pulse, was mediated by Na+-dependent transporters located exclusively at the basolateral membrane. One was independent of HCO3- and blocked by EIPA (probably NHE1) while the other was HCO3--dependent and blocked by H2DIDS (probably pNBC1). Changes in pH(i) following blockade of basolateral HCO3- accumulation confirmed that the cells achieve vectorial HCO3- secretion. Dose-dependent increases in HCO3- secretion were observed in response to stimulation of both secretin and VPAC receptors. ATP and UTP applied to the apical membrane stimulated HCO3- secretion but were inhibitory when applied to the basolateral membrane. HCO3- secretion in guinea-pig ducts and Capan-1 cell monolayers share many common features, suggesting that the latter is an excellent model for studies of human pancreatic HCO3- secretion.
机译：人的胰管分泌富含碳酸氢根的液体，但我们对分泌过程的了解主要基于动物模型的研究。我们的目的是确定人类导管细胞系中的HCO3-转运机制是否与先前在豚鼠胰导管中鉴定的机制相似。通过微荧光法在生长在可渗透滤膜上并装有BCECF的Capan-1细胞单层中测量细胞内pH。通过闭合蛋白的免疫定位评估上皮极化。通过RT-PCR评估关键电解质转运蛋白和受体的mRNA表达。生长在可渗透支持物上的Capan-1细胞形成了融合的，极化的单层，具有发达的紧密连接。由短时间的NH4 +脉冲诱导的从酸负荷中恢复pH（i）是由仅位于基底外侧膜的Na +依赖性转运蛋白介导的。一个独立于HCO3-，被EIPA（可能是NHE1）阻断，而另一个独立于HCO3-，被H2DIDS（可能是pNBC1）阻断。基底外侧HCO3-积累受阻后pH（i）的变化证实细胞实现了载体HCO3-的分泌。响应于促胰液素和VPAC受体的刺激，观察到HCO3-分泌的剂量依赖性增加。 ATP和UTP应用于根尖膜可刺激HCO3-分泌，但当应用于基底外侧膜时具有抑制作用。豚鼠导管和Capan-1细胞单层中的HCO3-分泌具有许多共同特征，这表明后者是研究人类胰腺HCO3-分泌的极好模型。

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