Hayashi M, Inagaki A, Novak I, Matsuda H
Department of Physiology, Kansai Medical University, 2-5-1 Shimmachi, Hirakata, 573-1010, Japan.
Medical Research Project, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
Pflugers Arch. 2016 Jul;468(7):1171-1181. doi: 10.1007/s00424-016-1806-9. Epub 2016 Mar 11.
Adenosine modulates a wide variety of biological processes via adenosine receptors. In the exocrine pancreas, adenosine regulates transepithelial anion secretion in duct cells and is considered to play a role in acini-to-duct signaling. To identify the functional adenosine receptors and Cl(-) channels important for anion secretion, we herein performed experiments on Capan-1, a human pancreatic duct cell line, using open-circuit Ussing chamber and gramicidin-perforated patch-clamp techniques. The luminal addition of adenosine increased the negative transepithelial potential difference (V te) in Capan-1 monolayers with a half-maximal effective concentration value of approximately 10 μM, which corresponded to the value obtained on whole-cell Cl(-) currents in Capan-1 single cells. The effects of adenosine on V te, an equivalent short-circuit current (I sc), and whole-cell Cl(-) currents were inhibited by CFTRinh-172, a cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel inhibitor. The adenosine A2B receptor agonist, BAY 60-6583, increased I sc and whole-cell Cl(-) currents through CFTR Cl(-) channels, whereas the A2A receptor agonist, CGS 21680, had negligible effects. The A2B receptor antagonist, PSB 603, inhibited the response of I sc to adenosine. Immunohistochemical analysis showed that the A2A and A2B receptors colocalized with Ezrin in the luminal membranes of Capan-1 monolayers and in rat pancreatic ducts. Adenosine elicited the whole-cell Cl(-) currents in guinea pig duct cells. These results demonstrate that luminal adenosine regulates anion secretion by activating CFTR Cl(-) channels via adenosine A2B receptors on the luminal membranes of Capan-1 cells. The present study endorses that purinergic signaling is important in the regulation of pancreatic secretion.
腺苷通过腺苷受体调节多种生物学过程。在外分泌胰腺中,腺苷调节导管细胞的跨上皮阴离子分泌,并被认为在腺泡到导管的信号传导中起作用。为了鉴定对阴离子分泌重要的功能性腺苷受体和Cl(-)通道,我们在此使用开路Ussing室和短杆菌肽穿孔膜片钳技术,对人胰腺导管细胞系Capan-1进行了实验。在Capan-1单层细胞中管腔添加腺苷可增加负跨上皮电位差(V te),其半数有效浓度值约为10 μM,这与在Capan-1单细胞中全细胞Cl(-)电流获得的值相对应。腺苷对V te、等效短路电流(I sc)和全细胞Cl(-)电流的作用被囊性纤维化跨膜电导调节因子(CFTR)Cl(-)通道抑制剂CFTRinh-172抑制。腺苷A2B受体激动剂BAY 60-6583通过CFTR Cl(-)通道增加I sc和全细胞Cl(-)电流,而A2A受体激动剂CGS 21680的作用可忽略不计。A2B受体拮抗剂PSB 603抑制I sc对腺苷的反应。免疫组织化学分析表明,A2A和A2B受体与埃兹蛋白共定位于Capan-1单层细胞的管腔膜和大鼠胰腺导管中。腺苷在豚鼠导管细胞中引发全细胞Cl(-)电流。这些结果表明,管腔腺苷通过激活Capan-1细胞管腔膜上的腺苷A2B受体来调节阴离子分泌。本研究支持嘌呤能信号在胰腺分泌调节中很重要。