Loss of CFTR-dependent anion and fluid secretion in the ducts of the exocrine pancreas is thought to contribute to the development of pancreatitis, but little is known about the impact of inflammation on ductal CFTR function. Here we used adult stem cell-derived cell cultures (organoids) obtained from porcine pancreas to evaluate the effects of pro-inflammatory cytokines on CFTR function. Organoids were cultured from porcine pancreas and used to prepare ductal epithelial monolayers. Monolayers were characterized by immunocytochemistry. Epithelial bicarbonate and chloride secretion, and the effect of IL-1β, IL-6, IFN-γ, and TNF-α on CFTR function was assessed by electrophysiology. Immunolocalization of ductal markers, including CFTR, keratin 7, and zonula occludens 1, demonstrated that organoid-derived cells formed a highly polarized epithelium. Stimulation by secretin or VIP triggered CFTR-dependent anion secretion across epithelial monolayers, whereas purinergic receptor stimulation by UTP, elicited CFTR-independent anion secretion. Most of the anion secretory response was attributable to bicarbonate transport. The combination of IL-1β, IL-6, IFN-γ, and TNF-α markedly enhanced CFTR expression and anion secretion across ductal epithelial monolayers, whereas these cytokines had little effect when tested separately. Although TNF-α triggered apoptotic signaling, epithelial barrier function was not significantly affected by cytokine exposure. Pro-inflammatory cytokines enhance CFTR-dependent anion secretion across pancreatic ductal epithelium. We propose that up-regulation of CFTR in the early stages of the inflammatory response, may serve to promote the removal of pathogenic stimuli from the ductal tree, and limit tissue injury.

中文翻译：

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促炎细胞因子刺激胰腺导管上皮中 CFTR 依赖性阴离子分泌

外分泌胰腺导管中 CFTR 依赖性阴离子和液体分泌的丧失被认为导致胰腺炎的发生，但人们对炎症对导管 CFTR 功能的影响知之甚少。在这里，我们使用从猪胰腺获得的成体干细胞衍生的细胞培养物（类器官）来评估促炎细胞因子对 CFTR 功能的影响。从猪胰腺中培养类器官并用于制备导管上皮单层。通过免疫细胞化学对单层细胞进行表征。通过电生理学评估上皮碳酸氢盐和氯化物的分泌，以及 IL-1β、IL-6、IFN-γ 和 TNF-α 对 CFTR 功能的影响。导管标记物（包括 CFTR、角蛋白 7 和闭合小带 1）的免疫定位表明，类器官来源的细胞形成了高度极化的上皮。促胰液素或 VIP 的刺激触发了跨上皮单层的 CFTR 依赖性阴离子分泌，而 UTP 的嘌呤能受体刺激则引发了 CFTR 独立的阴离子分泌。大多数阴离子分泌反应归因于碳酸氢根转运。IL-1β、IL-6、IFN-γ和TNF-α的组合显着增强了跨导管上皮单层的CFTR表达和阴离子分泌，而这些细胞因子在单独测试时几乎没有影响。尽管 TNF-α 触发细胞凋亡信号，但上皮屏障功能并未受到细胞因子暴露的显着影响。促炎细胞因子增强跨胰腺导管上皮的 CFTR 依赖性阴离子分泌。我们认为，在炎症反应的早期阶段 CFTR 的上调可能有助于促进导管树中致病刺激的消除，并限制组织损伤。