荷叶碱对野生型和突变型囊性纤维化跨膜电导调节因子氯离子通道的激活作用

中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (2): 138-143.

荷叶碱对野生型和突变型囊性纤维化跨膜电导调节因子氯离子通道的激活作用

林森, 侯曙光, 金伶伶, 于波, 杨红

辽宁师范大学生物制药中心, 大连 116029, 辽宁

收稿日期:2007-11-22 修回日期:2007-12-13 出版日期:2008-02-26 发布日期:2020-10-14
通讯作者: 杨红, 女, 教授, 博士生导师, 主要从事膜通道的结构与功能研究、药物高通量筛选及中药现代化研究。Tel:0411-84259113 　E-mail:hyang@nenu.edu.cn
作者简介:林森, 女, 硕士研究生, 主要从事膜通道的结构与功能研究及中药现代化研究。
基金资助:
国家自然科学基金资助项目(30470405, 30570864)

Natural compound nuciferine activates chloride channel of wild type and △F508 mutant cystic fibrosis transmembrane conductance regulator

LIN Sen, HOU Shu-guang, JIN Ling-ling, YU Bo, YANG Hong

Biopharmatheutical Center of Liaoning Normal University, Dalian 116029, Liaoning, China

Received:2007-11-22 Revised:2007-12-13 Online:2008-02-26 Published:2020-10-14

摘要/Abstract

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CFTRCl^-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CFTR 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。

关键词: 囊性纤维化跨膜电导调节因子, Cl^-转运, 荷叶碱, 激活剂

Abstract: 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:Nuciferine was identified as an effective activator of wild-type CFTR chloride channel by screening of 386 single compounds from Chinese medicinal herbs. The CFTR-stimulating activity is rapid and reversible. Nuciferine does not elevate cellular cAMP level and activates phophorylated CFTR, suggesting that it works by direct binding to CFTR molecule. Nuciferine 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 CFTR 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.

Key words: cystic fibrosis transmembrane conductance regulator, chloridion transport, nuciferine, potentiator

中图分类号:

R965.2

林森, 侯曙光, 金伶伶, 于波, 杨红. 荷叶碱对野生型和突变型囊性纤维化跨膜电导调节因子氯离子通道的激活作用[J]. 中国临床药理学与治疗学, 2008, 13(2): 138-143.

LIN Sen, HOU Shu-guang, JIN Ling-ling, YU Bo, YANG Hong. Natural compound nuciferine activates chloride channel of wild type and △F508 mutant cystic fibrosis transmembrane conductance regulator[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(2): 138-143.

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