法尼酯X受体下调肝型脂肪酸结合蛋白的表达.doc

法尼X受体 ( （第三军医大学：1基础部生物化学与分子生物学教研室，重庆 400038，2新桥医院血液科 重庆 400037） 　摘要 目的：法尼X受体( farnesoid X receptor，FXR )对肝型脂肪酸结合蛋白（liver fatty acid binding protein，L-FABP）表达的影响方法：μmol/L和100μmol/L的鹅脱氧胆酸（chenodesoxycholic acid，CDCA）以及1μmol/L和5μmol/L的GW4064分别处理人胎肝细胞L2和人肝癌细胞HepG2后，用逆转录-聚合酶链反应（RT-PCR）法检测FXR特异靶基因L-FABP的mRNA表达情况，再用Western 免疫印迹法检测L-FABP蛋白水平的变化）法尼X受体arnesoid X Receptor Down-regulates the Expression of Liver Fatty Acid Binding Protein DONG Jin-Yu1), LIU Hong2),PENG Jia-He1),ZHANG Yan1)，WANG Qiang1), LI Liang-Peng1),WANG Yong-Chao1),JIANG Yu1)( (1)Department of Biochemistry and Molecular Biology, Third Military Medical University, 2)Department of Clinical Laboratory ,Xinqiao Hospital , Chongqing 400037, China ) Abstract: Objective To investigate the effect of farnesoid X receptor (FXR) on the expression of liver fatty acid binding protein(L-FABP).Method: Human hepatoblastoma HepG2 cells and he- patocyte L02 cells were treated with FXR agonist chenodesoxycholic acid（CDCA） or GW4064. The L-FABP and the specific gene of FXR-- Small heterodimer partner （SHP） were detected by reverse transcription polymerase chain reaction (RT-PCR).The L-FABP protein level was determined by Western Blot .Result: After treated with CDCA or GW4064.SHP mRNA level in HepG2 and L02 was increased obviously，which suggested FXR was functional in those cells.The mRNA and protein levels of L-FABP decreased dramatically(p0.05) Conclusion: Farnesoid X receptor down-regulates the expression of liver Farnesoid X Receptor; liver fatty acid binding protein; metabolic syndrome；Hepatocytes 法尼X受体 [10,11]，提示 L-FABP在代谢性疾病的多个方面起重要作用。FXR和L-FABP肝脏中高表达，两者在脂代谢中均起重要作用，但FXR对L-FABP表达的影响尚未见报到，本实验通过研究FXR激动剂对表达，人肝癌细胞株HepG2由本室DMEM高糖培养基HyClone公司；CDCASigma公司；抗人克隆抗体美国Santa Cruz公司；辣根过氧化物酶标记抗二抗广州中杉公司 1.2 方法 1.2.1 细胞培养HepG2和L02细胞株含10%胎牛血清的DMEM高糖培养基37℃、5%的CO2温箱培养．2～4 d更换培养基次，待细胞到对数生长期时，细胞以×106/孔接种至孔板，每孔加含2%的培养ml，μmol/L和100μmol/L的CDCA以及1μmol/L和5μmol/L的GW4064分别处理L02和HepG2细胞，以溶解配体的溶剂DMSO(终浓度为1％o)作对照。24 h后按Trizol试剂操作说明提取细胞总RNA。． 1.PCR反应，