醋柴胡对HEK293细胞中MRP1蛋白摄取活性、蛋白表达及mRNA水平的影响

Influence of Vinegar-baked Radix Bupleuri on Protein Uptake Activity and mRNA Levels of MRP1 in HEK 293 Cells

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 explore the mechanism of Vinegar- baked Radix Bupleuri（VBRB） to MRP1 （multi -drug resistance protein 1 ） in HEK 293 cells. Methods ： Cell proliferation was analyzed by MTT assay. The uptake activity of MRP1 protein was measured by HPLC with colchicine as substrate. MRP1 protein and mRNA levels were detected by Western blot and RT - qPCR. Results：After treatment of VBRB （10 mg · mL- 1 ）for 48 hours, the uptake of colchicine and the mRNA level of MRPI were de- creased by 21% and 40%, respectively;while the protein expression of MRPt did not change. Conclusion：VBRB enhanced the ef- flux of MRP1 substrate colchicine,which activation might be not associated with MRP1 mRNA and protein expression.