{"title":"堪非醇通过 cAMP/PKA 信号通路激活 T84 细胞中氯化物的分泌和 CFTR 的表达","authors":"Janjira Thaweewattanodom, Chatsri Deachapunya, Sutthasinee Poonyachoti","doi":"10.3389/fphar.2024.1401273","DOIUrl":null,"url":null,"abstract":"Kaempferol is a flavonol identified as the most potent activator of chloride (Cl<jats:sup>−</jats:sup>) secretion among other flavonoids in airway epithelial cells. This study aimed to investigate the cellular mechanisms by which kaempferol stimulates Cl<jats:sup>−</jats:sup> secretion in the T84 human colon carcinoma cell line by Ussing chambers and voltage clamp technique. Bilateral addition of kaempferol (1–100 µM) increased short-circuit current (<jats:italic>I</jats:italic><jats:sub><jats:italic>sc</jats:italic></jats:sub>) in a concentration-dependent manner. Ion substitution of Cl<jats:sup>−</jats:sup> or CFTR inhibitors NPPB and glibenclamide or a Na<jats:sup>+</jats:sup>/K<jats:sup>+</jats:sup>/2Cl<jats:sup>−</jats:sup> cotransporter inhibitor bumetanide attenuated kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>sc</jats:italic></jats:sub> response. In permeabilized monolayers, selective channel inhibitors CFTRinh-172 and CaCCinh-A01 inhibited kaempferol-induced apical Cl<jats:sup>−</jats:sup> current (<jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub>), and K<jats:sup>+</jats:sup> blockers BaCl<jats:sub>2</jats:sub> and clotrimazole inhibited basolateral K<jats:sup>+</jats:sup> current (<jats:italic>I</jats:italic><jats:sub><jats:italic>Kb</jats:italic></jats:sub>). The kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub> showed no additive effects with forskolin or 8cpt-cAMP. The kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub> was mostly abolished by protein kinase A inhibitor H89, but not by tyrosine kinase inhibitors, AG490 and tyrphostin A23, or tyrosine phosphatase inhibitor vanadate. Treatment with kaempferol for 24 h increased the expression of CFTR protein as determined by the Western blot analysis. These results demonstrated that kaempferol activates Cl<jats:sup>−</jats:sup> secretion across T84 cells by activating the apical Cl<jats:sup>−</jats:sup> current and basolateral K<jats:sup>+</jats:sup> current. The mechanisms may involve the cAMP/PKA pathway and CFTR expression. Taken together, these findings reveal the beneficial effects of kaempferol to increase fluid secretion which can be used to treat constipation.","PeriodicalId":12491,"journal":{"name":"Frontiers in Pharmacology","volume":null,"pages":null},"PeriodicalIF":4.4000,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Kaempferol activates chloride secretion via the cAMP/PKA signaling pathway and expression of CFTR in T84 cells\",\"authors\":\"Janjira Thaweewattanodom, Chatsri Deachapunya, Sutthasinee Poonyachoti\",\"doi\":\"10.3389/fphar.2024.1401273\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Kaempferol is a flavonol identified as the most potent activator of chloride (Cl<jats:sup>−</jats:sup>) secretion among other flavonoids in airway epithelial cells. This study aimed to investigate the cellular mechanisms by which kaempferol stimulates Cl<jats:sup>−</jats:sup> secretion in the T84 human colon carcinoma cell line by Ussing chambers and voltage clamp technique. Bilateral addition of kaempferol (1–100 µM) increased short-circuit current (<jats:italic>I</jats:italic><jats:sub><jats:italic>sc</jats:italic></jats:sub>) in a concentration-dependent manner. Ion substitution of Cl<jats:sup>−</jats:sup> or CFTR inhibitors NPPB and glibenclamide or a Na<jats:sup>+</jats:sup>/K<jats:sup>+</jats:sup>/2Cl<jats:sup>−</jats:sup> cotransporter inhibitor bumetanide attenuated kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>sc</jats:italic></jats:sub> response. In permeabilized monolayers, selective channel inhibitors CFTRinh-172 and CaCCinh-A01 inhibited kaempferol-induced apical Cl<jats:sup>−</jats:sup> current (<jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub>), and K<jats:sup>+</jats:sup> blockers BaCl<jats:sub>2</jats:sub> and clotrimazole inhibited basolateral K<jats:sup>+</jats:sup> current (<jats:italic>I</jats:italic><jats:sub><jats:italic>Kb</jats:italic></jats:sub>). The kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub> showed no additive effects with forskolin or 8cpt-cAMP. The kaempferol-induced <jats:italic>I</jats:italic><jats:sub><jats:italic>Cl</jats:italic></jats:sub> was mostly abolished by protein kinase A inhibitor H89, but not by tyrosine kinase inhibitors, AG490 and tyrphostin A23, or tyrosine phosphatase inhibitor vanadate. Treatment with kaempferol for 24 h increased the expression of CFTR protein as determined by the Western blot analysis. These results demonstrated that kaempferol activates Cl<jats:sup>−</jats:sup> secretion across T84 cells by activating the apical Cl<jats:sup>−</jats:sup> current and basolateral K<jats:sup>+</jats:sup> current. The mechanisms may involve the cAMP/PKA pathway and CFTR expression. Taken together, these findings reveal the beneficial effects of kaempferol to increase fluid secretion which can be used to treat constipation.\",\"PeriodicalId\":12491,\"journal\":{\"name\":\"Frontiers in Pharmacology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2024-09-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Frontiers in Pharmacology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3389/fphar.2024.1401273\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PHARMACOLOGY & PHARMACY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Frontiers in Pharmacology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3389/fphar.2024.1401273","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PHARMACOLOGY & PHARMACY","Score":null,"Total":0}
Kaempferol activates chloride secretion via the cAMP/PKA signaling pathway and expression of CFTR in T84 cells
Kaempferol is a flavonol identified as the most potent activator of chloride (Cl−) secretion among other flavonoids in airway epithelial cells. This study aimed to investigate the cellular mechanisms by which kaempferol stimulates Cl− secretion in the T84 human colon carcinoma cell line by Ussing chambers and voltage clamp technique. Bilateral addition of kaempferol (1–100 µM) increased short-circuit current (Isc) in a concentration-dependent manner. Ion substitution of Cl− or CFTR inhibitors NPPB and glibenclamide or a Na+/K+/2Cl− cotransporter inhibitor bumetanide attenuated kaempferol-induced Isc response. In permeabilized monolayers, selective channel inhibitors CFTRinh-172 and CaCCinh-A01 inhibited kaempferol-induced apical Cl− current (ICl), and K+ blockers BaCl2 and clotrimazole inhibited basolateral K+ current (IKb). The kaempferol-induced ICl showed no additive effects with forskolin or 8cpt-cAMP. The kaempferol-induced ICl was mostly abolished by protein kinase A inhibitor H89, but not by tyrosine kinase inhibitors, AG490 and tyrphostin A23, or tyrosine phosphatase inhibitor vanadate. Treatment with kaempferol for 24 h increased the expression of CFTR protein as determined by the Western blot analysis. These results demonstrated that kaempferol activates Cl− secretion across T84 cells by activating the apical Cl− current and basolateral K+ current. The mechanisms may involve the cAMP/PKA pathway and CFTR expression. Taken together, these findings reveal the beneficial effects of kaempferol to increase fluid secretion which can be used to treat constipation.
期刊介绍:
Frontiers in Pharmacology is a leading journal in its field, publishing rigorously peer-reviewed research across disciplines, including basic and clinical pharmacology, medicinal chemistry, pharmacy and toxicology. Field Chief Editor Heike Wulff at UC Davis is supported by an outstanding Editorial Board of international researchers. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide.