Effect of the aqueous extract from Vinegar-baked Bupleuri Radix on the uptake of substrates by regulating various drug transporters in cells
Objective To determine the effect of aqueous extract from Vinegar-baked Bupleuri Radix(VBRB)on the uptake of drug transporters in BRL3A and HEK293 cells,and corresponding substrates,to elucidate the target for VBRB and provide data support for the action mechanism of VBRB meridian inducing and synergistic effect.Methods Sodium taurocholate,colchicine and rhodamine B were used as substrates for Ntcp/Oatp2,MRP1 and Pgp,respectively.The contents of substrates in the cells were determined by HPLC or flow cytometry.The protein concentration was measured by BCA method,and the uptake rate was calculated with the normalization of protein concentration.In addition,the effect of VBRB on the protein expression and mRNA level of drug transporters in the cells were analyzed by Western blot and RT-PCR.Results VBRB promoted the uptake of sodium taurocholate in BRL3A cells and increased the protein expression and mRNA levels of Ntcp and Oatp2.Similar to MRP1 inhibitor MK571,VBRB greatly increased the uptake of colchicine and decreased MRP1 protein expression in HEK293 cells when stimulated by glutathione.In addition,VBRB obviously promoted the uptake of Rhodamine B in Pgp-overexpression HEK293 cells,and inhibited the protein expression and mRNA level of Pgp.Conclusion VBRB enhances the uptake of multiple drug transporter substrates,which is related to its simultaneous activation of multiple influx transporters and inhibition of efflux transporters.
Vinegar-baked Bupleuri Radixmeridian inducing and synergistic effectNtcpOatp2MRP1Pgp
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