Electrophysiological Studies on the Evaluation of Absorption Enhancers in Caco-2 Cells Using a Microelectrode Technique
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概要
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We developed a microelectrode technique to characterize the electrophysiological properties in Caco-2 cells, and used it to determine the mechanisms of absorption enhancers. The action of absorption enhancers on the apical membrane of Caco-2 cells was estimated by measuring the apical membrane potential (V_m) with the microelectrode. The V_m value of Caco-2 cells in Hanks' balanced salt solution containing 0.5 mM K^+ was 18.9±0.8 mV (n=217), and the apical membrane resistance was 49.4±1.1 MΩ (n=160). In the electrophysiological study with absorption enhancers, laurylmaltoside (LM) markedly decreased the V_m value, while sodium glycocholate(NaGC) moderately reduced this value, and EDTA did not affect the value. These findings might be associated with their action sites, plasma membrane or tight junction in Caco-2 cell monolayers. In influx and transport studies with these absorption enhancers, LM enhanced the influx of furosemide, which is transported via both the transcellular and paracellular routes into Caco-2 cells, and enhanced its transport to the basolateral side of Caco-2 monolayers more than that of 5(6)-carboxyfluorescein (CF), a paracellular marker. EDTA did not increase the influx of furosemide, and enhanced the transport of furosemide and CF across Caco-2 cell monolayers to the same extent. In contrast, NaGC only slightly increased the influx of furosemide and did not enhance the transport of either furosemide or CF across the Caco-2 monolayers in this study. These findings were well correlated with the effects of these absorption enhancers on the electrophysiological parameters. Therefore, the microelectrode technique might be useful for evaluating the action of absorption enhancers in the plasma membrane at an intact cell level.
- 公益社団法人日本薬学会の論文
- 2000-06-01
著者
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YAMAMOTO Akira
Department of Biopharmaceutics, Kyoto Pharmaceutical University
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FUJITA Takuya
Department of Biopharmaceutics, Kyoto Pharmaceutical University
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KAMIYAMA Fumio
Department of Biopharmaceutics, Kyoto Pharmaceutical University
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Fujita T
Division Of Oral Cytology And Cell Biology Department Of Developmental And Reconstructive Medicine N
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FUJITA Takuya
Ritsumeikan University College of Information Science and Engeneering
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Quan Ying-shu
Department Of Biopharmaceutics Kyoto Pharmaceutical University:cosmed Pharmaceutical Co. Ltd.
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Yamamoto Akira
Dep. Of Biopharmaceutics Kyoto Pharmaceutical Univ.
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Kamiyama Fumio
京都大学再生医科学研究所
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Kodama Fusao
Cosmed Pharmaceutical Co. Ltd.
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Fujita T
Ritsumeikan University College Of Information Science And Engeneering
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Fujita Takuya
Department Of Basic Pharmaceutics Faculty Of Pharmaceutical Sciences Kyoto University
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Fujita T
Showa College Of Pharmaceutical Sciences
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Komada F
Department Of Pharmacy Kobe University School Of Medicine
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Fukushima Tohru
Department Of Pharmacology Faculty Of Pharmaceutical Sciences Setsunan University
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Yamamoto Akira
Department Of Biopharmaceutics Kyoto Pharmaceutical University
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Yamamoto Akira
Department Of Anatomy School Of Medicine University Of Tokushuma
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Yamamoto Akira
Department Of Anatomy School Of Medicine University Of Tokushima
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