芒果苷上调HepG2 细胞膜蛋白MRP3 和核受体PXR, CPF 表达.docVIP

芒果苷上调HepG2细胞膜MRP3和核受体PXR、CPF表达 张艳梅1,柴进1, 阳勇2，张婷1，陈文生1 （1第三军医大学西南医院全军消化病研究所，2重庆市中药研究院，重庆40038） [摘 要] 目的 体外细胞培养观察芒果苷（mangiferin）对多耐药相关蛋白MRP3（multidrg resistance-associate protein 3, ABCC3）和核受体PXR（pregnane X receptor）、CPF（CYP7A promoter-binding factor）表达的影响。方法 用芒果苷刺激HepG2细胞72h后，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转录与蛋白水平的表达变化。UDCA（ursodeoxycholic acid,UDCA）结果 芒果苷可显著刺激HepG2细胞膜转运蛋白MRP3，其作用强于UDCA。芒果苷也可上调核受体PXR、CPF的表达。结论 芒果苷刺激肝癌细胞HepG2细胞膜转运蛋白MRP3表达上调可能与核受体PXR、CPF途径相关。[关键词] 芒果苷;UDCA;MRP3;PXR;CPF Up-regulation of membrane transporter MRP3 and nuclear receptors PXR and CPF induced by mangiferin in HepG2 cells 1Zhang Yan-mei, 1CHAI Jin, 2YANG Yong, 1ZHANG Ting,1CHEN Wen-sheng (1Gastroenterology Institute, Third Military Medical University, Chongqing 400038; 2Chongqing Academy of Chinese Materia Medica,Chongqing 400065; China) Abstract:Objective To observe changes of expression of membrane transporter MRP3（multidrg resistance-associate protein 3,ABCC3）and nuclear receptor pregnane X receptor（PXR） and CYP7A promoter-binding factor（CPF）induced by mangiferin in HepG2 cells.Methods Both total RNA and total membrance or nuclear extracts were isolated from HepG2 cells after 72h treatment by mangiferin, or UDCA as an positive control, or DMSO as an negative control. The mRNA and protein levels of MRP3 ,PXR and CPF were determined by semiquantitative RT–PCR and western blot.Results The expression of membrane transporter MRP3 in HepG2 cells treated with mangiferin was dramatically increased compared with controls.And it was also more predominanced compared with positive controls. Nuclear receptors PXR and CPF was significantly up-regulated in HepG2 cells stimulated by mangiferin.Conclusion Up-regulation of membrane transporter MRP3 levels may be associated with nuclear receptor PXR and PXR in HepG2 cells treated by mangiferin. Keywords:mangiferin; UDCA;MRP3;PXR;CPF MRP3（multidrug resistance protein）MRP家族的成员广泛分布于人和啮齿类动物肝、胆道、肾脏、肠等组织，定位于极性化细胞的基底侧膜一种有机阴离子排泌泵，包含跨膜区和核甘酸结合域主要转运、GSH、硫酸盐