Involvement of cholesterol in the inhibitory effect of dimethyl‐β‐cyclodextrin on P‐glycoprotein and MRP2 function in Caco‐2 cells

FEBS Letters

Arima, Hidetoshi¹  Hirayama, Fumitoshi¹  Yunomae, Kiyokazu¹  Uekama, Kaneto¹ 

[1] Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan

关键词: Cyclodextrin;    Cholesterol;    P-glycoprotein;    Multidrug resistance-associated protein 2;    Caveolae;    Caco-2;    AP-to-BL;    apical to basolateral;    BCECF;    2′;    7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein;    BCECF-AM;    2′;    7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethylester;    BL-to-AP;    basolateral to apical;    CyDs;    cyclodextrins;    DS;    degree of substitution;    DM-β-CyD;    2;    6-di-O-methyl-β-cyclodextrin;    HP-α-CyD;    2-hydroxypropyl-α-cyclodextrin;    HP-β-CyD;    2-hydroxypropyl-β-cyclodextrin;    HP-γ-CyD;    2-hydroxypropyl-γ-cyclodextrin;    HPLC;    high performance liquid chromatography;    MRP2;    multidrug resistance-associated protein 2;    M-β-CyD;    methyl-β-cyclodextrin;    MBS;    MES-buffered saline;    MDR;    multidrug-resistant;    P app;    apparent permeability coefficient;    P-gp;    P-glycoprotein;    RT-PCR;    reverse transcription polymerase reaction;    SDS;    sodium dodecyl sulfate;    TEER;    transepithelial electric resistance;   

DOI  :  10.1016/S0014-5793(03)00059-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We compared the inhibitory effect of various cyclodextrins (CyDs) on P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2) function and examined the contribution of cholesterol to the inhibitory effect of 2,6-di-O-methyl-β-cyclodextrin (DM-β-CyD) on the efflux activity of the function in Caco-2 cell monolayers. Of various CyDs, DM-β-CyD significantly impaired the efflux activity of P-gp and MRP2. DM-β-CyD released P-gp and MRP2 from the monolayers in the apical side's transport buffer and decreased the extent of cholesterol as well as P-gp and MRP2 in caveolae of Caco-2 cell monolayers, but not caveolin and flotillin-1. On the other hand, DM-β-CyD did not change MDR1 and MRP2 mRNA levels. Therefore, these results suggest that the inhibitory effect of DM-β-CyD on P-gp and MRP2 function, at least in part, could be attributed to the release of these transporters from the apical membranes into the medium as secondary effects through cholesterol-depletion in caveolae after treatment of Caco-2 cell monolayers with DM-β-CyD.

【 授权许可】

Unknown