Resveratrol rescues cAMP-dependent anionic transport in the cystic fibrosis pancreatic cell line CFPAC1
Resveratrol rescues cAMP-dependent anionic transport in the cystic fibrosis pancreatic cell line CFPAC1
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DOI：
10.1111/j.1476-5381.2011.01289.x
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发表时间：
2011-06-01
影响因子：
7.3
通讯作者：
Planelles, Gabrielle
中科院分区：
文献类型：
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作者：
Hamdaoui, Nabila;Baudoin-Legros, Maryvonne;Planelles, Gabrielle
背景与目的囊性纤维化跨膜传导调节因子(CFTR)是上皮细胞质膜上cAMP依赖的氯离子通道，其突变是遗传性疾病囊性纤维化的原因。最常见的cftr突变是Phe(508)的缺失，这种突变蛋白(DelF508CFtr)不容易转移到质膜，在细胞内迅速降解。我们假设白藜芦醇(一种来自葡萄的天然多酚类植物雌激素化合物)处理上皮细胞2或18小时，可以调节CFTR的表达和定位。实验用白藜芦醇处理内源性表达CFTR(MDCK1和CAPAN1细胞)或delF508CFTR(来源于人支气管活检组织的CFPAC1和呼吸道上皮细胞)的细胞，用RT-PCR、Western印迹和免疫细胞化学方法评价白藜芦醇对CFTR和delF508CFTR表达和定位的影响。用荧光染料和卤化物敏感电极测量卤化物外排。结果白藜芦醇处理MDCK1细胞或CFPAC1细胞的免疫印迹实验中，白藜芦醇可促进CFTR的表达或成熟。间接免疫荧光实验显示，在CFPAC1细胞和人呼吸道上皮细胞中，delF508CFTR的定位向膜周区移动。白藜芦醇处理的CFPAC1细胞膜对卤化物的通透性呈cAMP依赖性增加，并可被选择性的cftr抑制剂抑制。结论与应用本实验结果表明，白藜芦醇可调节cftr的表达和定位，并能挽救依赖cAMP的氯的转运。
BACKGROUND AND PURPOSEThe cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-dependent chloride channel in the plasma membrane of epithelia whose mutation is the cause of the genetic disease cystic fibrosis (CF). The most frequent CFTR mutation is deletion of Phe(508) and this mutant protein (delF508CFTR) does not readily translocate to the plasma membrane and is rapidly degraded within the cell. We hypothesized that treating epithelial cells with resveratrol, a natural polyphenolic, phyto-ooestrogenic compound from grapes, could modulate both the expression and localization of CFTR.EXPERIMENTAL APPROACHCells endogenously expressing CFTR (MDCK1 and CAPAN1 cells) or delF508CFTR (CFPAC1 and airway epithelial cells, deriving from human bronchial biopsies) were treated with resveratrol for 2 or 18 h. The effect of this treatment on CFTR and delF508CFTR expression and localization was evaluated using RT-PCR, Western blot and immunocytochemistry. Halide efflux was measured with a fluorescent dye and with halide-sensitive electrodes. Production of interleukin-8 by these cells was assayed by ELISA.KEY RESULTSResveratrol treatment increased CFTR expression or maturation in immunoblotting experiments in MDCK1 cells or in CFPAC1 cells. Indirect immunofluorescence experiments showed a shift of delF508CFTR localization towards the (peri)-membrane area in CFPAC1 cells and in human airway epithelial cells. A cAMP-dependent increase in membrane permeability to halide was detected in resveratrol-treated CFPAC1 cells, and was inhibited by a selective inhibitor of CFTR.CONCLUSION AND IMPLICATIONSThese results show that resveratrol modulated CFTR expression and localization and could rescue cAMP-dependent chloride transport in delF508CFTR cells.