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目的体外HepG2细胞培养观察芒果苷(mangiferin)对多耐药相关蛋白3(multidrug resistance-associate protein 3,MRP3)和核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用芒果苷刺激HepG2细胞72 h后,分别抽提细胞RNA、膜蛋白及核蛋白,采用半定量RT-PCR和蛋白免疫印迹检测膜转运蛋白MRP3和核受体PXR、CPF在转录与蛋白水平的表达变化。熊脱氧胆酸(ursode-oxycholic acid,UDCA)处理的HepG2细胞作为阳性对照、DMSO处理细胞为阴性对照。结果芒果苷可显著刺激HepG2细胞膜转运蛋白MRP3的mRNA(比阴性对照组高3.0倍,P<0.05)和蛋白(比阴性对照组高3.3倍,P<0.05)表达,其作用强于UDCA。芒果苷也可明显上调核受体PXR[mRNA水平增高1.7倍(P<0.05),蛋白水平增高3.7倍(P<0.01)]、CPF[mRNA水平增高2.1倍(P<0.05),蛋白水平增高4.9倍(P<0.05)]的表达。结论芒果苷刺激肝癌细胞HepG2细胞膜转运蛋白MRP3的表达上调可能与核受体PXR、CPF途径相关。
Objective To investigate the effect of mangiferin on multidrug resistance-associate protein 3 (MRP3) and pregnane X receptor (PXR), CYP7A promoter binding factor (CYP7A promoter-binding factor, CPF) expression. Methods After stimulating HepG2 cells with mangiferin for 72 h, RNA, membrane protein and nuclear protein were extracted respectively. The mRNA and protein levels of MRP3 and PXR were measured by semi-quantitative RT-PCR and Western blot. Changes in expression. Ursodeoxycholic acid (UDCA) -treated HepG2 cells served as a positive control and DMSO-treated cells as a negative control. Results Mangiferin significantly stimulated mRNA expression of HepG2 cell membrane transporter MRP3 (3.0-fold higher than the negative control group, P <0.05) and protein (3.3-fold higher than the negative control group, P <0.05). Mangiferin also significantly up-regulated nuclear receptor PXR [mRNA level increased by 1.7-fold (P <0.05), protein level increased by 3.7-fold (P <0.01)], CPF [mRNA level increased by 2.1-fold 4.9-fold (P <0.05)]. Conclusion The up-regulation of MRP3 in HepG2 cells induced by mangiferin may be related to the nuclear receptor PXR and CPF.