β-eudesmol but not atractylodin exerts an inhibitory effect on CFTR-mediated chloride transport in human intestinal epithelial cells
β-eudesmol but not atractylodin exerts an inhibitory effect on CFTR-mediated chloride transport in human intestinal epithelial cells
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β-eudesmol 但不是苍术素对人肠上皮细胞中 CFTR 介导的氯离子转运产生抑制作用
DOI：
10.1016/j.biopha.2021.112030
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发表时间：
2021
影响因子：
7.5
通讯作者：
Muanprasat, Chatchai
中科院分区：
文献类型：
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作者：
Tharabenjasin, Phuntila;Ferraris, Ronaldo P.;Choowongkomon, Kiattawee;Pongkorpsakol, Pawin;Worakajit, Nichakorn;Sawasvirojwong, Sutthipong;Pabalan, Noel;Na-Bangchang, Kesara;Muanprasat, Chatchai
东方草药具有两种生物活性成分，β-苦楝酚（BE）和白术素（AT），已被用作治疗胃肠道疾病的药物。没有科学证据报道它们的止泻作用和基础机制。因此，我们旨在研究这两种化合物的体外抗分泌活性。采用Ussing chamber法观察BE和AT对人肠上皮（T84）细胞camp诱导的cl -分泌的抑制作用。加入间接和直接囊性纤维化跨膜电导调节剂（CFTR）氯离子通道激活剂后测定短路电流（ISC）和根尖cl -电流（ICl-）。MTT法测定细胞毒性。通过硅分子对接分析研究了蛋白质与配体的相互作用。在T84细胞单层上，经上皮检测，BE可降低camp介导的CFTRinh-172抑制的Cl -分泌，但不依赖于AT浓度（ic50为~1.05µM）。使用不同的CFTR激活剂，BE对CFTR介导的icl抑制的效力并没有改变，这表明CFTR通道活性位点被直接阻断。BE预处理完全阻止camp诱导的1 -。此外，浓度高达200µM （24 h）的BE对T84细胞活力没有影响。在硅中，研究表明BE可以在核苷酸结合域（NBD） 2区域的atp结合口袋上与CFTR的去磷酸化结构对接。这些发现为BE的抗分泌作用包括抑制CFTR功能提供了第一个证据。BE是治疗或预防肠道Cl-高分泌引起的腹泻的一个有希望的候选物
Oriental herbal medicine with the two bioactive constituents, β-eudesmol (BE) and atractylodin (AT), has been used as a remedy for gastrointestinal disorders. There was no scientific evidence reporting their antidiarrheal effect and underpinning mechanisms. Therefore, we aimed to investigate the anti-secretory activity of these two compoundsin vitro.The inhibitory effect of BE and AT on cAMP-induced Cl-secretion was evaluated by Ussing chamber in human intestinal epithelial (T84) cells. Short-circuit current (ISC) and apical Cl-current (ICl-) were measured after adding indirect and direct cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activator. MTT assay was used to determine cellular cytotoxicity. Protein-ligand interaction was investigated byin silicomolecular docking analysis. BE, but not AT concentration-dependently (IC50of ~1.05 µM) reduced cAMP-mediated, CFTRinh-172 inhibitable Cl−secretion as determined by transepithelialISCacross a monolayer of T84 cells. Potency of CFTR-mediatedICl-inhibition by BE did not change with the use of different CFTR activators suggesting a direct blockage of the channel active site(s). Pretreatment with BE completely prevented cAMP-inducedICl-. Furthermore, BE at concentrations up to 200 µM (24 h) had no effect on T84 cell viability.In silicostudies indicated that BE could best dock onto dephosphorylated structure of CFTR at ATP-binding pockets in nucleotide-binding domain (NBD) 2 region. These findings provide the first evidence for the anti-secretory effect of BE involving inhibition of CFTR function. BE represents a promising candidate for the therapeutic or prophylactic intervention of diarrhea resulted from intestinal hypersecretion of Cl-