中文摘要：

目的：从天然单体化合物中筛选能够激活囊性纤维化跨膜电导调节因子（CFTR）Cl^-通道转运活性的激活剂。方法：利用稳定表达人CFTR和对卤族元素碘离子高度敏感的荧光绿蛋白突变体EYFP—H148Q的Fischer大鼠甲状腺上皮细胞为筛选模型，测定386种中药单体化合物对CFTR介导的I^-内流速度的影响。结果：发现生物碱类化合物荷叶碱对野生型和△F508突变型CFTR Cl^-通道具有激活作用，而对G551D突变型CFTRCl^-通道无激活作用。荷叶碱对野生型和△F508突变型CFTR Cl^-通道的激活作用具有作用迅速、可逆的特点，并且依赖于细胞内cAMP水平。荷叶碱不提高细胞内cAMP水平，当CFTR磷酸化程度达到最大时仍能进一步激活CFTR Cl^-通道激活作用，表明它可能是通过与CFYR直接结合而发挥作用的。结论：首次发现荷叶碱对野生型和△F508突变型CFTR Cl^-通道的激活作用，该天然化合物将在阐明CFR活性机制及作为先导化合物开发与CFTR有关的疾病治疗药物等方面具有重要用途。

英文摘要：

AIM： To identify CFTR Cl^- channel gating potentiators from natural compounds. METHODS： A stably transfected Fischer thyroid epithelial （FRT） cell line co-expressing human CFTR and a green fluorescent protein mutant with ultra-high halide sensitivity （YFP-H148Q） was used to measure CFTR- mediated iodide influx rate stimulated by the 386 natural compounds. RESULTS： Nueifefine was identified as an effective activator of wild-type CFTR ebloride ehannel by screening of 386 single eompounds from Chinese medieinal herbs. The CFTR-stimulating activity is rapid and reversible. Nueiferine does not elevate eellular eAMP level and activates phophorylated CFTR, suggesting that it works by direct binding to CFTR mol-ecule. Nucifefine can also potentiate the Cl^- transport of △F508 mutant CFTR. CONCLUSION： Nuciferine was identified as an effective CFTR chloride channel activator. Nuciferine may be useful for probing CFFR channel gating mechanisms and as a lead compound to develop pharmacological therapy of CFTR-related disease such as cystic fibrosis, idiopathic chronic pancreatiti, keratoconjunctivitis sicca and habitual constipation.