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目的以Caco-2细胞模型筛选妥舒沙星、斯帕沙星、辛伐他汀是否为P-糖蛋白(P-gp)的底物。方法以Caco-2细胞为模型,以酮康唑、兰索拉唑为对照,用流式细胞仪测定妥舒沙星、斯帕沙星、辛伐他汀对Caco-2细胞表面的P-gp转运罗丹明-123的影响,经筛选得到可能为P-gp底物的药物后,再用药物转运实验,验证其是否为P-gp的底物。结果妥舒沙星和斯帕沙星存在明显抑制P-gp向细胞外转运罗丹明-123的作用;在加入P-gp抑制剂(酮康唑)后,妥舒沙星的PappBL→AP/PappAP→BL由10.03±0.47降至0.93±0.19,斯帕沙星PappBL→AP/PappAP→BL由2.54±0.12降至1.02±0.04。结论流式细胞仪联用罗丹明-123可简单有效地从多种药物中筛选出可能是P-gp底物的药物(如妥舒沙星和斯帕沙星)及转运机制等。
Objective To screen whether tosufloxacin, sparfloxacin and simvastatin are substrates of P-glycoprotein (P-gp) by Caco-2 cell model. Methods Caco-2 cells as a model, ketoconazole, lansoprazole as a control, flow cytometry tosufloxacin, sparfloxacin, simvastatin on Caco-2 cell surface P-gp After transfection of Rhodamine-123, we found that P-gp could be the substrate of P-gp after screening. Results tosufloxacin and sparfloxacin significantly inhibited the P-gp translocation to rhodamine-123. After addition of P-gp inhibitor (ketoconazole), tosufloxacin PappBL → AP / PappAP → BL decreased from 10.03 ± 0.47 to 0.93 ± 0.19, while that of sparfloxacin decreased from 2.54 ± 0.12 to 1.02 ± 0.04 PappBL → AP / PappAP → BL. Conclusion Flow cytometry combined Rhodamine-123 can be simply and effectively selected from a variety of drugs may be P-gp substrate drugs (such as tosufloxacin and sparfloxacin) and transport mechanisms.