大豆苷原对成骨细胞MC3T3-E1甾体激素受体辅激活因子1表达的调节作用及其机制  

Effects of daidzein on steroid receptor coactivator-1 expression in MC3T3-E1 cells and the mechanism

作　　者：王立芳[1] 王金凤[1] 金慰芳[1] 王洪复[1] 张绍芬[2] 高建军[1] 

机构地区：[1]复旦大学放射医学研究所骨代谢研究室,上海200032 [2]复旦大学妇产科医院妇科,上海200011

出　　处：《中西医结合学报》2011年第11期1248-1253,共6页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30872756)

摘　　要：目的:研究大豆苷原对成骨细胞MC3T3-E1甾体激素受体辅激活因子1(steroid receptorcoactivator-1,SRC-1)和核受体辅抑制因子(nuclear receptor corepressor,NcoR)表达的调节作用及其机制。方法:体外培养MC3T3-E1细胞于含2%胎牛血清(fetal bovine serum,FBS)的α最低必须培养基(α-minimal essential medium,α-MEM)中,给予不同浓度大豆苷原或10^(-8)mol/L的雌二醇作用3 d后收获细胞,采用蛋白质印迹法观察SRC-1和NcoR蛋白的表达水平。分别应用雌激素受体(estrogen receptor,ER)拮抗剂ICI182780(Faslodex,ICI,10^(-7) mol/L)和雌激素受体α(estrogen receptorα,ERα)特异性拮抗剂(methyl-piperidino-pyrazole,MPP,10^(-6) mol/L)干预,观察大豆苷原对细胞SRC-1和NcoR蛋白表达水平的影响。结果:大豆苷原可上调MC3T3-E1细胞SRC-1的表达,10^(-7)mol/L和10^(-5)mol/L大豆苷原组SRC-1的蛋白水平分别增加至对照组的2.5倍(P<0.05)和2倍(P<0.05)。各剂量组NcoR表达水平与对照组比较差异无统计学意义。雌二醇组SRC-1水平与对照组比较未见明显变化,而其NcoR表达水平较对照组下调35%(P<0.05)。ICI182780可拮抗大豆苷原对SRC-1的上调作用。在ICI182780作用下,各剂量组SRC-1蛋白水平与对照组比较差异无统计学意义;MPP干预下,10^(-7)mol/L和10^(-5) mol/L大豆苷原组SRC-1的蛋白水平分别为对照组的1.8倍(P<0.05)和2.4倍(P<0.05)。结论:大豆苷原可增加成骨细胞SRC-1的蛋白表达水平,提高细胞SRC-1/NcoR比值。ERβ参与了大豆苷原对SRC-1的调节过程。成骨细胞内SRC-1蛋白的增加和SRC-1/NcoR比值的提高是大豆苷原促进成骨细胞分化的机制之一。Objective： To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 （SRC-1） and nuclear receptor corepressor （NcoR） in MC3T3-E1 osteoblastic cells. Metheds. MC3T3-E1 cells were cultured in e-minimal essential medium （α-MEM） containing 2% fetal bovine serum and treated with various concentrations of daidzein （10^-9 , 10^-7 and 10-s mol/L） or 17β-estradiol at 10^-8 mol/L for 3 d. The protein levels of SRC-1 and NcoR in MC3T3-E1 cells were determined by Western blotting. Estrogen receptor （ER） antagonist ICI182780 at 10^- 7 mol/L or specific ERe antagonist methyl-piperidino- pyrazole （MPP） at 10 6 mol/L were used to block the corresponding receptors, and then MC3T3-E1 cells were treated with daidzein at 10^-7 mol/L or 10-5 mol/L for 3 d. SRC-1 and NcoR protein levels were detected by Western blotting. Results： The protein levels of SRC-1 increased by 2.5 fold （P〈0.05） and 2 fold （P〈0.05） by 10-7 and 10-s mol/L of daidzein respectively, while the NcoR levels were not significantly altered. 1713-Estradiol at dose of 10-8 mol/L did not affect the expression of SRC-1 but decreased NcoR protein expression by 35% （P〈0. 05）. Compared with the control, daidzein at 10^-7 and 10^-5 mol/L did not increase SRC-1 expression when ERs were blocked by antagonist ICI182780. Daidzein at 10^-7 and 10^-5 mol/L up-regulated SRC-1 by 1.8 fold （P〈0. 05） and 2.4 fold （P〈0.05） respectively while ERe was blocked by MPP. Conclusion= Daidzein increases protein level of SRC-1 and the ratio of SRC-1/NcoR. ERI3, instead of ERe, participates in the action of daidzein in regulating SRC-1 expression. Up-regulation of SRC-1 and increase of SRC-1/NcoR are part of the mechanism of the estrogenic effect of daidzein in improving osteogenesis.

关 键 词：大豆苷原 甾体激素受体辅激活因子1 核受体辅抑制因子 成骨细胞 受体 雌激素 

分 类 号：R285.5[医药卫生—中药学]