摘要
目的考察醋柴胡水提液(VBRB)对大鼠肝细胞BRL3A以及人胚肾细胞HEK293内多种药物转运蛋白的影响以及对相应底物的摄取,进一步确定醋柴胡引经增效的作用靶点,为全面解析醋柴胡引经增效的作用机制提供数据支撑。方法分别选用牛磺胆酸钠为Ntcp、Oatp2的共同底物,秋水仙碱、罗丹明B作为MRP1以及Pgp的底物,通过HPLC法或流式细胞术测定细胞内各底物含量;同时采用BCA法测定蛋白浓度,按蛋白质浓度归一化法计算摄取量;另分别采用Western blot法和RT-PCR法分析VBRB对细胞中的多种转运蛋白的蛋白表达以及mRNA水平的影响。结果VBRB促进BRL3A细胞对牛磺胆酸钠摄取,并升高摄入型转运蛋白Ntcp和Oatp2的蛋白及mRNA水平。与MRP1抑制剂MK571类似,经谷胱甘肽刺激后的HEK293细胞,VBRB显著增加细胞对秋水仙碱摄取并降低MRP1蛋白表达;同时,VBRB显著促进Pgp过表达的HEK293细胞对罗丹明B摄取,并显著抑制Pgp蛋白表达和mRNA水平。结论醋柴胡可增强细胞对多种底物的摄取,与其同时激活细胞内多种摄取型药物转运蛋白和抑制外排型药物转运蛋白活性有关。
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 Pgpoverexpression 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.
作者
冯丽敏
张娴
赵瑞芝
FENG Li-min;ZHANG Xian;ZHAO Rui-zhi(State Key Laboratory of Chinese Medicine for Dampness Syndrome,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510006;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome,Guangzhou 510120)
出处
《中南药学》
CAS
2024年第7期1807-1811,共5页
Central South Pharmacy
基金
国家自然科学基金项目(No.81073063,No.82173981,No.81573612)。
关键词
醋柴胡
引经增效
NTCP
Oatp2
MRP1
PGP
Vinegar-baked Bupleuri Radix
meridian inducing and synergistic effect
Ntcp
Oatp2
MRP1
Pgp
