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

基金项目：国家自然科学基金资助项目 CQ CSTC 2007BA5030） Supporyed by the National Natural Science Foundation of China and CQ CSTC 2007BA5030) 作者简介：张艳梅（1983-），女，重庆市人，硕士研究生，主要从事肝脏膜转运蛋白结构与功能方面。 HYPERLINK mailto:E@mail:zhangyanmei1202@163.com Email:zhangyanmei1202@163.com 联系电话通讯作者：陈文生，电话：（023，Email:wenshengchen@ 芒果苷上调HepG2细胞膜蛋白MRP3和核受体 PXR、CPF表达 张艳梅1,柴进1, 阳勇2，张婷1，罗维早2，钟国跃2，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400038） [摘 要] 目的 体外HepG2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，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。UDCA（ursodeoxycholic 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和蛋白表达，其作用强于UDCA。芒果苷也可明显上调核受体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的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, 2LUO Wei-zhao, 2ZHONG Guo-yue ,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 protein 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 dr