Objectives: To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism.Methods: The apparent permeability coefficient(Papp) for the transport of vin...Objectives: To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism.Methods: The apparent permeability coefficient(Papp) for the transport of vincristine through the membrane of MDCK-MDR1 cells was used as an indicator of the effect of bergapten on vincristine transport.Molecular docking was employed to predict the binding force between bergapten and P-glycoprotein(Pgp). The effects of bergapten on P-gp function and P-gp ATPase activity were determined by rhodamine123(Rho123) accumulation and activity analysis, respectively. 1,6-Diphenyl-1,3,5-hexatriene(DPH) was used to study the effects of bergapten on membrane fluidity, and Western blotting and quantitative realtime PCR assays were performed to analyze the effect of bergapten on the protein and mRNA expression of P-gp, respectively. These experiments clarified the effects of bergapten on the transport of vincristine and allowed exploration of the possible mechanism underlying the effects of bergapten.Results: The results showed that bergapten could inhibit the transport of vincristine in MDCK-MDR1 cells,and the binding force between bergapten and P-gp was weaker. Bergapten could reduce the accumulation of Rh123 in MDCK-MDR1 cells, increase the membrane fluidity, and upregulate P-gp protein and mRNA expression but it had no effect on P-gp ATPase activity.Conclusions: Overall, we concluded that the possible mechanism through which bergapten inhibits vincristine transport was related to the bergapten-mediated upregulation of P-gp protein and mRNA expression, membrane fluidity or P-gp enzyme activity.展开更多
基金supported by a project of the National Natural Science Foundation of China(Grant No.81303237)the Training Project of Young Scientists in Jiangxi Province(No.20153BCB23019)+1 种基金China Postdoctoral Science Foundation(2016M590606)the National Natural Science Foundation of Jiangxi Province(20161ACB21020)
文摘Objectives: To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism.Methods: The apparent permeability coefficient(Papp) for the transport of vincristine through the membrane of MDCK-MDR1 cells was used as an indicator of the effect of bergapten on vincristine transport.Molecular docking was employed to predict the binding force between bergapten and P-glycoprotein(Pgp). The effects of bergapten on P-gp function and P-gp ATPase activity were determined by rhodamine123(Rho123) accumulation and activity analysis, respectively. 1,6-Diphenyl-1,3,5-hexatriene(DPH) was used to study the effects of bergapten on membrane fluidity, and Western blotting and quantitative realtime PCR assays were performed to analyze the effect of bergapten on the protein and mRNA expression of P-gp, respectively. These experiments clarified the effects of bergapten on the transport of vincristine and allowed exploration of the possible mechanism underlying the effects of bergapten.Results: The results showed that bergapten could inhibit the transport of vincristine in MDCK-MDR1 cells,and the binding force between bergapten and P-gp was weaker. Bergapten could reduce the accumulation of Rh123 in MDCK-MDR1 cells, increase the membrane fluidity, and upregulate P-gp protein and mRNA expression but it had no effect on P-gp ATPase activity.Conclusions: Overall, we concluded that the possible mechanism through which bergapten inhibits vincristine transport was related to the bergapten-mediated upregulation of P-gp protein and mRNA expression, membrane fluidity or P-gp enzyme activity.
