CAJ | 학술논문

目的探讨P-糖蛋白抑制剂对两性霉素B(AmB)血脑屏障转运的影响.方法应用小鼠脑血管内皮细胞建立体外血脑屏障模型,选取维拉帕米作为P-糖蛋白抑制剂,AmB作为受试底物,经细胞毒性实验选定合适的温育时间及药物浓度,分别进行不同时间点的AmB细胞摄取实验及不同维拉帕米浓度的AmB细胞摄取实验.组间均数比较采用单因素方差分析,均数两两比较采用Bonferroni检验.结果 AmB的血脑屏障转运随时间延长不断积累,与同一时间点对照组相比,抑制剂组在90 min(t=6.753,P=0.001)、120 min(t=3.574,P=0.016)、150 min(t=4.759,P=0.005)处AmB细胞摄取水平明显升高,其余时间点两组间差异无统计学意义.与阴性对照组相比,AmB细胞摄取水平在不同浓度维拉帕米组(2、5、10、50、75和100μmol/L)均有显著提高(P=0.000、0.014、0.000、0.014、0.000、0.000),且随维拉帕米浓度升高增加幅度更明显.结论 P-糖蛋白抑制剂维拉帕米可提高脑毛细血管内皮细胞对AmB的摄取作用,提示P-糖蛋白参与血脑屏障AmB的转运.
목적 탐토P-당단백억제제대량성매소B(AmB)혈뇌병장전운적영향.방법 응용소서뇌혈관내피세포건입체외혈뇌병장모형,선취유랍파미작위P-당단백억제제,AmB작위수시저물,경세포독성실험선정합괄적온육시간급약물농도,분별진행불동시간점적AmB세포섭취실험급불동유랍파미농도적AmB세포섭취실험.조간균수비교채용단인소방차분석,균수량량비교채용Bonferroni검험.결과 AmB적혈뇌병장전운수시간연장불단적루,여동일시간점대조조상비,억제제조재90 min(t=6.753,P=0.001)、120 min(t=3.574,P=0.016)、150 min(t=4.759,P=0.005)처AmB세포섭취수평명현승고,기여시간점량조간차이무통계학의의.여음성대조조상비,AmB세포섭취수평재불동농도유랍파미조(2、5、10、50、75화100μmol/L)균유현저제고(P=0.000、0.014、0.000、0.014、0.000、0.000),차수유랍파미농도승고증가폭도경명현.결론 P-당단백억제제유랍파미가제고뇌모세혈관내피세포대AmB적섭취작용,제시P-당단백삼여혈뇌병장AmB적전운.
Objective To determine the influence of P-glycoprotein (P-gp) inhibitor on the blood brain barrier (BBB) transport of amphotericin B (AmB)..Methods An in-vitro BBB model was established with brain capillary endothelia cells (BCEC). AmB was chosen as the test drug and verapamil was chosen as the inhibitor of P-gp.Cellular uptake of AmB at different time points and with series of verapamil concentrations were performed respectively after the determination of appropriate incubation time and drug dosage by the cytotoxicity assay. The AmB concentrations of series of samples were detected using high performance liquid chromatography (HPLC) method. One-way ANOVA analysis and Bonferroni test were used for data analysis.Results The cellular transport of AmB was accumulated as the time prolonged.The inhibitor group had a significant higher cellular uptake levelsof AmBat the time point of 90 min (t=6.753,P=0.001),120 min (t=3.574,P=0.016) and 150 min (t=4.759,P=0.005) as compared with the control group.The AmB cellular uptake level increased significantly when BCEC were incubated with verapamil of 2 μmol/L (P=0.000),5 μmol/L (P=0.014),10 μmol/L (P=0.000),50 μmol/L (P=0.014),75 μmol/L (P=0.000) and 100 tμmol/L (P=0.000),respectively,compared with the control group.Conclusion The P-gp inhibitor verapamil can enhance the cellular uptake of AmB which indicates that P-gp is involved in the BBP transport of AmB.