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目的:研究醋柴胡水提液对人胚肾细胞HEK293中MRP1影响的作用机制。方法:MTT法观察醋柴胡水提液对HEK293细胞增殖的影响;蛋白摄取活性实验以秋水仙碱作底物,采用高效液相色谱(HPLC)法检测胞内底物含量;采用Western blot法检测蛋白表达水平;荧光定量RT-PCR法检测mRNA水平。结果:与对照组相比,醋柴胡水提液(10mg·mL-1)作用48 h后,细胞对秋水仙碱的摄取率降低了21%,差异有统计学意义(P<0.01);相应的MRP1 mRNA水平降低了40%,差异有统计学意义(P<0.05);而蛋白表达则无变化。结论:醋柴胡能增加胞内MRP1转运底物秋水仙碱的外排,但可能并非通过改变MRP1 mRNA和蛋白表达水平而实现。
Objective: To study the mechanism of action of water extract of vinegar on MRP1 in human embryonic kidney cell line HEK293. Methods: MTT assay was used to observe the effect of aqueous extract of Aceh Bupleurum on the proliferation of HEK293 cells. The activity of protein uptake was determined by using colchicine as the substrate. The contents of intracellular substrate were determined by high performance liquid chromatography (HPLC) The protein expression level was detected by fluorescence quantitative RT-PCR. Results: Compared with the control group, the uptake rate of colchicine was decreased by 21% (P <0.01) after treated with water extract of vinegar (10 mg · mL -1) for 48 h. The corresponding MRP1 mRNA level decreased by 40%, the difference was statistically significant (P <0.05); while the protein expression did not change. Conclusion: Bupleurum vinegar can increase the efflux of intracellular MRP1 transporter colchicine, but may not be achieved by changing MRP1 mRNA and protein expression level.