摘要
目的研究治疗剂量下4种抗结核药物(异烟肼、左氧氟沙星、乙胺丁醇、吡嗪酰胺)对Caco-2细胞上P-糖蛋白功能、表达及MDR1 mRNA表达的影响,从而解释联合抗痨治疗中不同组合的合理性。方法采用流式细胞仪测定细胞内罗丹明-123的浓度,考察药物对P-糖蛋白功能的影响,流式细胞术分析药物对Caco-2细胞上P-糖蛋白表达的影响,实时荧光定量PCR技术分析药物对Caco-2细胞MDR1基因mRNA水平表达的影响。结果含药培养20 d后,异烟肼和乙胺丁醇减少了罗丹明-123在Caco-2细胞内的蓄积(P<0.05),为诱导作用;异烟肼、乙胺丁醇均上调了Caco-2细胞上P-糖蛋白的表达(P<0.05),其P-糖蛋白表达量分别为阴性对照组的3.5和3.8倍;同时上调了Caco-2细胞上MDR1 mRNA的表达(P<0.05),其MDR1 mRNA的表达量分别为阴性对照组的11.5和11倍。左氧氟沙星增加了罗丹明-123在Caco-2细胞内的蓄积(P<0.05),为抑制作用;下调了Caco-2细胞上P-糖蛋白和MDR1 mRNA的表达(P<0.05),其P-糖蛋白和MDR1 mRNA表达量分别为阴性对照组的50%和32%。而吡嗪酰胺与P-糖蛋白无明显相互作用。结论治疗剂量的异烟肼和乙胺丁醇为P-糖蛋白的诱导剂,左氧氟沙星为P-糖蛋白的抑制剂,而吡嗪酰胺对P-糖蛋白功能和表达无明显影响。
OBJECTIVE To study the effects of four anti-tuberculosis drugs on the function and expression of P-glycoprotein and the MDR1 mRNA expression,for explaining the rationality of different combination in anti-tuberculosis chemotherapy.METHODS The effect of the drugs on P-glycoprotein function was analyzed using Rh-123 assay.The flow cytometry was used to determine the intracellular Rh-123 concentration and the expression of P-glycoprotein in Caco-2 cells.Real-time fluorescent quantitative polymerase chain reaction was used to measure the expression of MDR1 gene mRNA in Caco-2 cells.RESULTS After the intervention for 20 d,the therapeutic doses of isoniazid and ethambutol decreased the accumulation of rhodamine123 in Caco-2 cells significantly compared with that of negative control group（P〈0.05）.The therapeutic doses of isoniazid and ethambutol both up-regulated the cellular P-glycoprotein protein and MDR1 mRNA expression levels（P〈0.05）.Compared with the controls,the total quantity of P-glycoprotein were 3.5 and 3.8 folds higher than that of controls,and the total levels of MDR1 mRNA expression were 11.5 and 11 folds higher than that of controls,respectively.Therapeutic doses of levofloxacin increased the accumulation of rhodamine123 in Caco-2 cells significantly higher than that of negative control group（P〈0.05）.The therapeutic doses of levofloxacin down-regulated the cellular P-glycoprotein protein and MDR1 mRNA expression levels（P〈0.05）.Compared with the controls,the total levels of P-glycoprotein and MDR1 mRNA expression were 50% and 32% of controls,respectively.Pyrazinamide showed no significant interaction with P-glycoprotein.CONCLUSION Therapeutic doses of isoniazid and ethambutol might be inducer of P-glycoprotein,levofloxacin might be inhibitors of p-glycoprotein,and pyrazinamide showed no significant interaction with P-glycoprotein.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2011年第1期48-53,共6页
Chinese Pharmaceutical Journal
