川陈皮素对囊性纤维化跨膜传导调节因子的激活作用.docVIP

  川陈皮素对囊性纤维化跨膜传导调节因子 的激活作用# 杨爽，于波，张耀方，王雪，杨红** 5 10 15 20 25 30 （辽宁师范大学生命科学学院，辽宁省生物技术与药物研发重点实验室，大连 116033） 摘要：目的：研究川陈皮素（ 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。方法：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，测定川陈皮素对 CFTR Cl－通道活性的影响。 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 陈皮素能够有效地刺激小鼠气管粘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。 关键词：细胞生物学；囊性纤维化跨膜传导因子；川陈皮素；激活剂 中图分类号：R96 The activation effect of nobiletin on cystic fibrosis transmenbrance conductance regulator chloride channel Yang Shuang, YU Bo, Zhang Yaofang, Wang Xue, Yang Hong (Liaoning Provincial Key Labroatory of Biotechnology and Drug Discovery,Liaoning Normal University,Dalian,116033) Abstract: Aim: Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator （ CFTR ） chloride channel activity. Method: CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Result: Nobiletin potently activated CFTR chaloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTRinh-172 could completely reverse the effect. Primary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may present a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis. Key words: cell biology; CFTR; nobiletin; activator 35 0 引言 囊性纤维化跨膜传导调节因子（cystic fibrosis transmembrane conductance regulator, CFTR）是一种 cAMP 依赖的氯离子通道蛋白，也是迄今为止发现的唯一作为通道的 ATP 结合蛋白家族（ATP-binding cassette, ABC）成员[1]。CFTR 由 5 个功能结构域组成：2 个由  40 6 个连续串联的跨膜区组成的跨膜结构域（membrane-sp