inducing resuscitation with herbal aromatics is important to modulate the brain intake of drugs in traditional Chinese medicine,but limited information has been available on the mechanism of action.The MDCK-MDRl monol...inducing resuscitation with herbal aromatics is important to modulate the brain intake of drugs in traditional Chinese medicine,but limited information has been available on the mechanism of action.The MDCK-MDRl monolayer is an excellent in vitro cell model to use as a tool to study blood brain barrier（BBB） screening.In this study,we established MDCK-MDR1 cell line by stable transfection and investigated the effects of several important herbal aromatics on BBB permeability.The characterization experiment demonstrated the MDCK-MDRl used in this study was valid.In a transport study,we found several herbal aromatics increased the permeability of fluorescein isothiocyanate-labeled dextran 4kDa（FD4） and inhibited efflux of Rhodaminel23（Rhol23）.These results demonstrated that herbal aromatics enhanced the BBB permeation of drugs by both inhibition of P-gp and opening of the BBB tight junction,thus providing new insights for understanding the mechanisms of aromatic compounds＇ BBB permeability.展开更多
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Major New Medicine Project in Mega-projects of Science Research of China(Grant No.2009ZX09502-006)National Natural Science Foundation of China(Grant No.20971008)
文摘inducing resuscitation with herbal aromatics is important to modulate the brain intake of drugs in traditional Chinese medicine,but limited information has been available on the mechanism of action.The MDCK-MDRl monolayer is an excellent in vitro cell model to use as a tool to study blood brain barrier（BBB） screening.In this study,we established MDCK-MDR1 cell line by stable transfection and investigated the effects of several important herbal aromatics on BBB permeability.The characterization experiment demonstrated the MDCK-MDRl used in this study was valid.In a transport study,we found several herbal aromatics increased the permeability of fluorescein isothiocyanate-labeled dextran 4kDa（FD4） and inhibited efflux of Rhodaminel23（Rhol23）.These results demonstrated that herbal aromatics enhanced the BBB permeation of drugs by both inhibition of P-gp and opening of the BBB tight junction,thus providing new insights for understanding the mechanisms of aromatic compounds＇ BBB permeability.
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.
