Influence Of Angelica Sinensis Extracts And Bergapten On Voriconazole Or Itraconazole Transport In Vitro

OBJECTIVEThrough the establishment of Caco-2 cell model, predict and select drug that may occur drug-drug interactions form Angelica sinensis extracts and their furocoumarins;In order to study the Influence of Angelica sinensis extracts and bergapten on voriconazole (VOR) or itraconazole (ITR) transport in vitro. Initially investigate the drug-drug interaction mechanisms of Angelica sinensis extracts and bergapten with VOR or ITR, which may provide the foundation for the theoretical basis of clinical medication safety.METHODS1 Caco-2 cell monolayer model and validationIn order to study the Influence of Angelica sinensis extracts and bergapten on VOR or ITR transport in vitro, while the use of Caco-2 cell culture method, constructed the Caco-2 cell monolayers in vitro. Caco-2 cells were identified by morphology, immunofluorescence, fluorescent yellow, propranolol permeability and transepithelial electrical resistance (TEER) etc.2 The cytotoxicity of AE, AW, BGT, VOR and ITR on Caco-2MTT (diphenyltetrazolium bromide) assay was used to find the non-cytotoxicity dosage of Angelica sinensis alcohol extract (AE), Angelica sinensis aqueous extract (AW), bergapton (BGT), VOR, ITR in Caco-2 cells. The dosage at which the survival rate of the cell is above 90% was considered as the highest non-cytotoxic dosage.3 Influence of Angelica sinensis extracts and bergapten on voriconazole transport in vitroHPLC method for determination of VOR was developed and validated.In order to investigate the two-way transmembrane transport, we have established Caco-2 cell monolayer model to compare VOR with co-administration of only (control), AE, AW, BGT. The amount of the bi-direction transportation of VOR was analysed with HPLC, and the apparent permeability coefficient (Papp) and excretion ratio (ER) were obtained.Cultured in the conventional Caco-2 cells, we examined the Caco-2 cell uptake experiments to compare VOR with co-administration of only (control), AE, AW, BGT. HPLC medthod for determination of the concentration of VOR was used.4 Influence of Angelica sinensis extracts and bergapten on itraconazole transport in vitroHPLC method for determination of ITR was developed and validated.In order to investigate the two-way transmembrane transport, we have established in vitro Caco-2 cell monolayer model to compare ITR with co-administration of only (control), AE, AW, BGT. The amount of the bi-direction transportation of ITR was analysed with HPLC, and the apparent permeability coefficient (Papp) and excretion ratio (ER) were obtained.Cultured in the conventional Caco-2 cells, we examined the Caco-2 cell uptake experiments to compare ITR with co-administration of only (control), AE, AW, BGT. HPLC medthod for determination of the concentration of ITR was used.RESULTS1 Caco-2 cell monolayer model and validationCaco-2 cells grew well under normal conditions, by cell morphology, immunofluorescence, fluorescent yellow and propranolol permeability and TEER of the verification, Which can meet the Caco-2 cell model requirements. These Angelica sinensis extract and its active constituents can be used to investigate the interaction of VOR and ITR.2 The cytotoxicity of AE, AW, BGT, VOR and ITR on Caco-224-hour MTT toxicity test results, AE, AW, BGT, VOR and ITR on Caco-2 cells, the maximum non-toxic concentrations were250μg-m-1, 250μg-mL-1, 30μg-mL-1,40μg-mL-1 and 50μg-mL-1, respectively. Were selected 20μg-mL-1 ITR and VOR,5,20μg-mL-1 BGT,100. 200μg-mL-1 AE and AW for next research.3 Influence of Angelica sinensis extracts and bergapten on voriconazole transport in vitroCaco-2 cell monolayers transport results show that:compared with the VOR only control group, VOR+AW group were significantly increased efflux, VOR+AE group was significantly reduced efflux, VOR+BGT group were no significant difference. ER=Pappba/Pappab≥1.71, it prompted VOR may be active transport in Caco-2 cells and may be subject to other carriers.In Caco-2 cell uptake results show that:compared with the VOR only control group, VOR+BGT and VOR+AE group had no effect on uptake, VOR+AW group were significantly reduced the uptake, differences between doses had no effect on uptake. Description:In the Caco-2 cells, AW can significantly reduce the uptake of VOR, while had no significant for BGT and AE, and is not combined with the impact of dose.4 Influence of Angelica sinensis extracts and bergapten on itraconazole transport in vitroThe transport Experimental of ITR results show that:compared with the ITR only control group, ITR+AW group were significantly increased efflux; ITR+BGT and ITR+AE group were significantly decreased efflux, and inhibition of efflux Rate:ITR+BGT>ITR+AE.BL-> AP direction of the apparent permeability is several times higher than the AP-> BL direction. ER=Pappba/Pappab≥1.03, it prompted ITR is active transport by carrier in transit transport. In Caco-2 cell uptake results show that:compared with the ITR only control group, ITR+BGT group increased significantly uptake of ITR in Caco-2 cells, ITR+AW group was significantly reduced uptake of ITR in Caco-2 cells, ITR+AE group showed no significant difference; differences between doses had no effect on uptake. Description:BGT can increase uptake of ITR, AW can decrease uptake of ITR, while AE had no significant for AE, and is not combined with the impact of dose.CONCLUSION1. Angelica sinensis alcohol extract, to some extent, have inhibited the efflux of voriconazole and itraconazole.2. Angelica sinensis aqueous extract, to some extent, have induced the efflux of voriconazole and itraconazole.3. Bergapton have inhibited the efflux of itraconazole, to some extent.