利用Caco-2细胞单层模型评价P-糖蛋白对安妥沙星转运的影响

Evaluation of P-gp on the transport of antofloxacin by using Caco-2 cell monolayer model

目的评价P-糖蛋白抑制剂酮康唑对安妥沙星转运的影响。方法利用人源结肠腺癌系Caco-2细胞单层模型对安妥沙星进行双向转运,采用高效液相色谱法对安妥沙星进行定量分析,计算其表观渗透系数(Papp)。结果加入酮康唑后,低浓度安妥沙星(100μmol/L)Papp(B-A)/Papp(A-B)由4.12降至1.23(P<0.01),高浓度安妥沙星(500μmol/L)Papp(B-A)/Papp(A-B)由3.54降至1.22(P<0.01)。结论安妥沙星在Caco-2细胞单层模型中的转运机制可能以主动转运为主,P-糖蛋白参与其从细胞基底侧向管腔面的分泌。

Objective To evaluate the effect of P-gp inhibitor ketoconazole on the transport of antofloxacin by using human colon adenocarcinoma line Caco-2 cell monolayer model. Methods The concentrations of antofloxacin collected from the transport assay were determined by high performance liquid chromatography and the transport parameters such as apparent permeability coefficient[Papp （A-B） and Papp （B-A）] and the ratio of Papp （B-A） versus Papp （A-B） were calculated and compared when the antofloxacin was used sololy and co-used with ketoconazole. Results When ketoconazole was present, the Papp （B-A）/Papp （A-B） of antofloxacin decreased from 4.12 to 1.23 and 3.54 to 1.22 at the concentration of 100μmol/L and 500μmol/L, respectively. And Papp （B-A） were also decreased obviously at each concentration level, from 4.23±0.27 to 2.39±0.70 and 8.49±0.89 to 5.03±1.20. Conclusion The reasults indicated that active transport might play a major role in the transport of antofloxacin in Caco-2 cell monolayer model and P-gp involved in the secretion of antofloxacin from basolateral to apical.