克拉霉素对Caco-2细胞转运他克莫司的影响及机制研究

Effect of Clarithromycin on Tacrolimus Transport in Caco-2 Cells and Its Mechanism

关键词:  他克莫司  克拉霉素  Caco-2细胞  转运

Key words:  tacrolimus  clarithromycin  Caco-2 cells  transport

摘要:

目的 探讨克拉霉素对Caco-2细胞转运他克莫司的影响及机制。方法 用化学发光免疫分析法测定他克莫司药物浓度；建立Caco-2细胞单层模型，计算表观渗透系数，比较不同浓度他克莫司在Caco-2细胞单层中转运及不同浓度克拉霉素抑制他克莫司的转运。结果 他克莫司浓度为20，40和80 μg·mL^-1时吸收渗透系数P_appAP-BL分别为（2.47±0.09）×10^-6，（3.91±0.17）×10^-6和（4.49±0.16）×10^-6 cm·s^-1；分泌渗透系数P_appBL-AP分别为（6.05±0.21）×10^-6，（9.86±0.70）×10^-6和（11.75±0.28）×10^-6 cm·s^-1；外流比（apparent permeability ratio，PDR）分别为2.45±0.03，2.52±0.12和2.62±0.11；加入不同浓度的克拉霉素（15，30，60 μg·mL^-1）后，分泌渗透系数显著降低，而吸收系数影响不大，PDR随着克拉霉素浓度增加显著降低。结论 Caco-2细胞外排转运体可能参与了他克莫司的转运，克拉霉素合用他克莫司可显著影响他克莫司的吸收。

Abstract:

OBJECTIVE To explore the effect and the mechanism of clarithromycin on the transport of tacrolimus in Caco-2 cells. METHODS The concentration of tacrolimus was determined by chemiluminescence immunoassay. The single layer model of Caco-2 cells was established, the apparent permeability coefficient was calculated, and to compare the transport of tacrolimus in Caco-2 cell monolayer at different concentrations and the inhibition of tacrolimus transport by clarithromycin at different concentrations. RESULTS When tacrolimus concentration was 20, 40 and 80 μg·mL^-1, the absorption coefficient P_appAP-BL was (2.47±0.09)×10^-6, (3.91±0.17)×10^-6 and (4.49±0.16)×10^-6cm·s^-1, respectively, and the secretory coefficient P_appBL-AP was (6.05±0.21)×10^-6, (9.86±0.70)×10^-6and (11.75±0.28)×10^-6cm·s^-1 respectively; the apparent permeability ratio(PDR) was 2.45±0.03, 2.52±0.12 and 2.62±0.11 respectively. After adding different concentrations of clarithromycin(15, 30, 60 μg·mL^-1), the secretory permeability coefficient decreased significantly, but the absorption coefficient was not affected, and the PDR decreased significantly with the increase of clarithromycin concentration. CONCLUSION The extracellular transporters of Caco-2 may be involved in the transport of tacrolimus and the absorption tacrolimus could be apparently affected by combination of clarithromycin.