Department of Pharmacology and Pharmacotherapy, University of Szeged, Szeged, Hungary.
Gut. 2011 Mar;60(3):361-9. doi: 10.1136/gut.2010.214213. Epub 2010 Oct 12.
Acute pancreatitis is among the few inflammatory diseases for which no specific pharmacological treatment is available. It has previously been shown that bile acids alter pancreatic ductal secretion and these effects are probably involved in the pathogenesis of bile-induced pancreatitis.
To understand the mechanism responsible for bile-induced hypersecretion and, in particular, to identify the molecular target for bile acids in native pancreatic duct epithelial cells (PDECs).
Patch clamp recordings and spectrofluorimetry were used to measure whole cell currents and rates of HCO₃⁻ secretion, respectively, from isolated guinea pig pancreatic ducts. Expression of ion channels and receptors was investigated by immunohistochemistry/immunofluorescence of intact pancreatic tissue.
Exposing PDECs to chenodeoxycholate (CDC, 100 μM) reversibly increased whole cell K(+) currents and hyperpolarised cell membrane potential. Bile acid-stimulated K(+) currents were inhibited by Ba²(+) (2 mM), iberiotoxin (100 nM), and suppressed by strong intracellular Ca²(+) buffering. Luminally applied iberiotoxin also blocked CDC-stimulated HCO₃⁻secretion from microperfused ducts; however, the inhibitor did not influence the stimulatory effect of secretin, carbachol or luminally applied ATP. The specific large-conductance Ca²(+)-activated potassium (BK) channel activator, NS11021, induced a similar increase in HCO₃⁻secretion to CDC. Immunohistochemical analysis showed strong BK channel protein expression on the apical membrane of PDECs, while the G-protein-coupled bile acid receptor-1 was not detected in PDECs, but was present in acinar cells.
It was shown for the first time that BK channels (i) are expressed at the apical membrane of guinea pig PDECs; (ii) have a crucial role in regulating HCO₃⁻ secretion and (iii) are also essential for the bile acid-induced hypersecretion and, therefore, underlie the response of the pancreas to this noxious agent.
急性胰腺炎是少数几种尚无特定药物治疗的炎症性疾病之一。先前的研究表明,胆汁酸会改变胰腺导管的分泌,这些作用可能与胆源性胰腺炎的发病机制有关。
了解胆汁诱导高分泌的机制,特别是确定天然胰腺导管上皮细胞(PDECs)中胆汁酸的分子靶点。
使用膜片钳记录和光谱荧光法分别测量分离的豚鼠胰腺导管的全细胞电流和 HCO₃⁻分泌率。通过对完整胰腺组织的免疫组织化学/免疫荧光染色来研究离子通道和受体的表达。
将 PDECs 暴露于鹅脱氧胆酸(CDC,100 μM)可使全细胞 K⁺电流可逆性增加,并使细胞膜电位超极化。胆酸盐刺激的 K⁺电流被 Ba²⁺(2 mM)、iberiotoxin(100 nM)抑制,并被强烈的细胞内 Ca²⁺缓冲抑制。腔内应用的 iveriotoxin 也阻断了微灌注导管中 CDC 刺激的 HCO₃⁻分泌;然而,该抑制剂并不影响促胰液素、乙酰胆碱或腔内应用 ATP 的刺激作用。特异性大电导钙激活钾(BK)通道激活剂 NS11021 诱导与 CDC 相似的 HCO₃⁻分泌增加。免疫组织化学分析显示,BK 通道蛋白在 PDECs 的顶端膜上表达强烈,而 G 蛋白偶联胆汁酸受体-1 未在 PDECs 中检测到,但在腺泡细胞中存在。
首次表明 BK 通道(i)在豚鼠 PDECs 的顶端膜上表达;(ii)在调节 HCO₃⁻分泌中起关键作用;(iii)也是胆汁酸诱导高分泌所必需的,因此是胰腺对这种有毒物质的反应的基础。