Analysis of Carrier-Mediated Transport Systems in Drug Absorption from Oral Mucosa

口腔粘膜药物吸收中载体介导的转运系统分析

基本信息

批准号：
10672041
负责人：
KIMURA Toshikiro
金额：
$ 2.05万
依托单位：
OKAYAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672041/
关键词：
human oral mucosal cells D-glucose dipeptide cultured stratified cell layer active transport facilitated diffusion transporter

项目摘要

The in vitro uptake study was performed using the isolated cells of human oral mucosa, buccal and the dorsum of tongue, to investigate the mechanisms of glucose and oligopeptide uptake. The uptake of D-glucose was much larger in cells of the dorsum of tongue than in buccal cells and was inhibited more extensively by 2-deoxy-D-glucose, a substrate of facilitative glucose transporters, than by α-methyl-D-glucoside, a specific substrate of SGLT1, suggesting the larger contribution of facilitative transporter than NaィイD1+ィエD1/glucose cotransporter. Furthermore, from the results of inhibition studies by the several sugar analogues including maltose and D-mannose, GLUT1 and/or GLUT3 were suggested to take part in the glucose uptake by oral mucosa. Therefore, we have attempted to confirm the expression of glucose transporters on the oral mucosa by employing Western blotting. As a result, it was suggested that SGLT1, GLUT1, GLUT2 and GLUT3 are expressed in the epithelial cells of human oral mucosa. The result of the uptake study of glycylsarcosine, a model dipeptide, by isolated oral cells showed the presence of carrier-mediated transport systems for oligopeptides in human oral mucosal cells. The transport of D-glucose and glycylsarcosine across stratified cell layer of cultured human oral mucosal cells was also examined, and the results showed that the transporters function not only for the uptake of these substrates by the mucosal cells but also for the transport across the stratified mucosal cell layers.

利用人口腔粘膜、颊部和舌背的分离细胞进行体外摄取研究，以研究葡萄糖和寡肽的摄取机制。舌背细胞对D-葡萄糖的摄取远高于舌背细胞。在口腔细胞中，其主要受到促进葡萄糖转运蛋白底物 2-脱氧-D-葡萄糖的抑制，而不是 α-甲基-D-葡萄糖苷（葡萄糖转运蛋白的特定底物）的抑制。 SGLT1，表明促进转运蛋白比 NaiD1+D1/葡萄糖协同转运蛋白的贡献更大。此外，根据包括麦芽糖和 D-甘露糖在内的几种糖类似物的抑制研究结果，表明 GLUT1 和/或 GLUT3 参与葡萄糖。因此，我们尝试通过采用蛋白质印迹法来确认口腔粘膜上的葡萄糖转运蛋白的表达。结果表明，SGLT1、GLUT1、GLUT2和GLUT3在人口腔粘膜上皮细胞中表达。分离的口腔细胞对甘氨酰肌氨酸（一种模型二肽）的摄取研究结果表明存在载体介导的转运系统。用于人口腔粘膜细胞中的寡肽 D-葡萄糖和甘氨酰肌氨酸穿过培养的人口腔粘膜细胞的分层细胞层的转运。还进行了检查，结果表明转运蛋白不仅具有粘膜细胞摄取这些底物的功能，而且还具有跨分层粘膜细胞层的转运功能。