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本实验利用荧光淬灭实验和膜片钳技术在稳定表达人CFTR和荧光绿蛋白突变体EYFP/H148Q的Fischer大鼠甲状腺上皮细胞(Fischer rat thyroid,FRT)上,测定川陈皮素(nobiletin)对囊性纤维化跨膜传导因子(cystic fibrosis transmembrane conductance regulator,CFTR)氯离子通道的激活作用。结果发现,川陈皮素以剂量依赖的方式激活CFTR氯离子通道的Cl-转运活性,且这种活性是快速、可逆的,并能够被CFTR特异性抑制剂CFTRinh-172完全抑制。初步的分子机制研究表明,川陈皮素是以与CFTR直接作用来激活通道活性的。进一步的研究结果显示,川陈皮素能够有效刺激小鼠气管黏膜下腺液体分泌速度。因此,川陈皮素可能发展成为治疗包括支气管扩张在内的CFTR相关疾病的先导药物。
In this study, Fischer rat thyroid (FST) thyroid epithelial cells (FST) stably expressing human CFTR and fluorescent green protein mutant EYFP / H148Q were assayed by fluorescence quenching and patch clamp techniques, and nobiletin Cystic fibrosis transmembrane conductance regulator (cystic fibrosis transmembrane conductance regulator, CFTR) chloride ion channel activation. As a result, it was found that nobiletin inhibited the Cl-transport activity of the CFTR chloride channel in a dose-dependent manner, and this activity was rapid and reversible and completely inhibited by CFTRinh-172, a CFTR-specific inhibitor. Preliminary molecular mechanism studies have shown that nobiletin is a direct interaction with CFTR to activate channel activity. Further research results show that nobiletin can effectively stimulate the tracheal submucosal fluid secretion in mice. Therefore, nobiletin may develop into a leading drug for the treatment of CFTR-related diseases including bronchiectasis.