Department of Pharmacology, Institute of Gastroenterology, Brain Korea 21 Project for Medical Science, Seoul, Korea.
Gastroenterology. 2010 Aug;139(2):620-31. doi: 10.1053/j.gastro.2010.04.004. Epub 2010 Apr 14.
BACKGROUND & AIMS: Pancreatic bicarbonate (HCO3-) secretion is important for a healthy pancreas as well as digestive physiology. However, how human pancreatic duct cells secrete copious amounts of HCO3- has long been a puzzle. Here, we report that a dynamic increase in the cystic fibrosis transmembrane conductance regulator (CFTR) HCO3- permeability by intracellular Cl- concentration ([Cl-]i)-sensitive mechanisms plays a pivotal role in pancreatic HCO3- secretion.
The role of [Cl-]i-sensitive kinases in CFTR-mediated HCO3- transport was examined in heterologous expression systems, PANC1 human pancreatic duct cells, and human and guinea pig pancreatic tissues using an integrated molecular and physiologic approach.
In human pancreatic tissues, CFTR-positive duct cells abundantly expressed with-no-lysine (WNK1) kinase, oxidative stress-responsive kinase 1 (OSR1), and sterile 20/SPS1-related proline/alanine-rich kinase (SPAK), which are known to be activated by low [Cl-]i. Interestingly, CFTR activation rapidly decreased [Cl-]i in response to luminal Cl- depletion in polarized PANC1 human pancreatic duct cells. Notably, the WNK1-mediated OSR1 and SPAK activation by low [Cl-]i strongly increased CFTR HCO3- permeability in CFTR-transfected HEK 293T, PANC1, and guinea pig pancreatic duct cells, making CFTR primarily an HCO3- channel, which is essential for the secretion of pancreatic juice containing HCO3- at a concentration greater than 140 mmol/L. In contrast, OSR1 and SPAK activation inhibited CFTR-dependent Cl-/HCO3- exchange activity that may reabsorb HCO3- from the high HCO3--containing pancreatic juice.
These results indicate that the [Cl-]i-sensitive activation of the WNK1-OSR1/SPAK pathway is the molecular switch to generate HCO3--rich fluid in the human pancreatic duct.
胰腺碳酸氢盐(HCO3-)的分泌对健康的胰腺和消化生理都很重要。然而,人类胰腺导管细胞如何大量分泌 HCO3- 长期以来一直是个谜。在这里,我们报告细胞内氯离子浓度([Cl-]i)敏感机制使囊性纤维化跨膜电导调节因子(CFTR)HCO3-通透性动态增加,在胰腺 HCO3-分泌中起着关键作用。
采用异源表达系统、PANC1 人胰腺导管细胞和人及豚鼠胰腺组织,综合分子和生理方法,研究[Cl-]i 敏感激酶在 CFTR 介导的 HCO3-转运中的作用。
在人胰腺组织中,CFTR 阳性导管细胞丰富表达无赖氨酸(WNK1)激酶、氧化应激反应激酶 1(OSR1)和无菌 20/SPS1 相关脯氨酸/丙氨酸丰富激酶(SPAK),已知这些激酶可被低 [Cl-]i 激活。有趣的是,CFTR 激活后可迅速降低细胞内氯离子浓度,以响应极化的 PANC1 人胰腺导管细胞中腔侧 Cl-的耗竭。值得注意的是,低 [Cl-]i 介导的 WNK1 对 OSR1 和 SPAK 的激活可显著增加 CFTR 转染的 HEK 293T、PANC1 和豚鼠胰腺导管细胞的 CFTR HCO3-通透性,使 CFTR 主要成为 HCO3-通道,这对分泌 HCO3-浓度大于 140mmol/L 的胰腺液是必需的。相反,OSR1 和 SPAK 的激活抑制 CFTR 依赖性 Cl-/HCO3-交换活性,可能会从富含 HCO3-的胰腺液中重吸收 HCO3-。
这些结果表明,WNK1-OSR1/SPAK 通路的 [Cl-]i 敏感激活是产生人胰腺导管中富含 HCO3-液体的分子开关。