Department of Otolaryngology, University of Alabama at Birmingham, Birmingham, AL.
Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL.
Int Forum Allergy Rhinol. 2019 Jan;9(1):100-105. doi: 10.1002/alr.22202. Epub 2018 Aug 27.
Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene result in defective Cl transport and cause chronic bacterial infections in the upper and lower airways of cystic fibrosis (CF) patients. Ivacaftor is a CFTR potentiator that improves Cl transport in CF patients with at least 1 copy of the G551D mutation. Resveratrol is also a potent CFTR potentiator that increases determinants of mucociliary transport. The objective of this study is to determine whether resveratrol and ivacaftor improve Cl secretion in G551D CFTR over either agent alone.
Fisher rat thyroid cells (FRT) transfected with G551D CFTR and human sinonasal epithelial cells (HSNE) containing the CFTR G551D mutation were subjected to pharmacologic manipulation of transepithelial ion transport in Ussing chambers. Activity was further evaluated using whole-cell patch clamp methods in G551D FRT cells.
In G551D FRT cells, resveratrol (100 μM) and ivacaftor (10 μM) significantly increased Cl transport (change in short-circuit current, δI = μA/cm ) compared with single-agent and dimethylsulfoxide vehicle controls (resveratrol + ivacaftor 4.97 ± 0.57 vs ivacaftor 0.74 ± 0.12 vs resveratrol 2.96 ± 0.52 vs control 0.74 ± 0.12; p < 0.001). Maximal Cl secretion (20 μM forskolin) was also significantly enhanced (p < 0.0001). Activity was confirmed in G551D HSNE (resveratrol + ivacaftor 4.48 ± 0.39 vs ivacaftor 1.05 ± 0.11 vs. resveratrol 0.84 ± 0.3 vs control, 0.0 ± 0.02; p < 0.001), and whole-cell patch clamp analysis in G551D FRT cells (resveratrol + ivacaftor -2535 ± 179.3 pA vs ivacaftor -1408.9 ± 101.3 pA vs resveratrol; -766.2 ± 71.2 pA; p < 0.0001).
Additive improvement in G551D CFTR-mediated Cl secretion suggests that resveratrol could enhance ivacaftor therapy in these patients and improve CF-related rhinosinusitis.
囊性纤维化跨膜电导调节因子 (CFTR) 基因突变导致氯离子转运缺陷，使囊性纤维化 (CF) 患者的上、下呼吸道发生慢性细菌感染。依伐卡托是一种 CFTR 增强剂，可改善至少携带 1 个 G551D 突变的 CF 患者的氯离子转运。白藜芦醇也是一种有效的 CFTR 增强剂，可增加黏液纤毛运输的决定因素。本研究旨在确定白藜芦醇和依伐卡托联合用药是否比单独用药更能改善 G551D CFTR 的氯离子分泌。
在含有 CFTR G551D 突变的 Fisher 大鼠甲状腺细胞 (FRT) 和人鼻黏膜上皮细胞 (HSNE) 中进行跨上皮离子转运的药物处理，并在 Ussing 室中进行研究。在 G551D FRT 细胞中进一步使用全细胞膜片钳方法评估活性。
在 G551D FRT 细胞中，与单药和二甲亚砜载体对照相比，白藜芦醇 (100 μM) 和依伐卡托 (10 μM) 显著增加氯离子转运 (短电路电流变化，δI=μA/cm )(白藜芦醇+依伐卡托 4.97±0.57 比依伐卡托 0.74±0.12 比白藜芦醇 2.96±0.52 比对照 0.74±0.12;p<0.001)。20 μM forskolin 诱导的氯离子分泌也显著增强(p<0.0001)。在 G551D HSNE 中也得到了证实(白藜芦醇+依伐卡托 4.48±0.39 比依伐卡托 1.05±0.11 比白藜芦醇 0.84±0.3 比对照 0.0±0.02;p<0.001)，并在 G551D FRT 细胞中进行全细胞膜片钳分析(白藜芦醇+依伐卡托 -2535±179.3 pA 比依伐卡托 -1408.9±101.3 pA 比白藜芦醇 -766.2±71.2 pA;p<0.0001)。
G551D CFTR 介导的氯离子分泌的累加改善表明，白藜芦醇可增强这些患者的依伐卡托治疗，并改善 CF 相关的鼻-鼻窦炎。
