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目的 研究槲皮素对大鼠原代肝细胞中Ⅰ型血红素氧化酶(HO-1)诱导的分子机制.方法 二步胶原酶技术分离培养大鼠原代肝细胞.不同剂量槲皮素作用大鼠原代肝细胞不同时间后,用RT-PCR方法检测HO-1的mRNA表达;50μmol/L槲皮素分别与10μmol/L PD98059、10μ mol/L SB203580、10 μ mol/L SP600125、1μmol/L Wormannin共孵育原代大鼠肝细胞12h后,用RT-PCR、Western blot法检测HO-1 mRNA和核因子E2相关因子2(Nrf2)蛋白表达水平的变化.对数据进行单因素方差分析、Bonferroni t检验.结果 大鼠原代肝细胞经25 ～ 200μmol/L槲皮素处理12h,或用50μmol/L槲皮素处理4～ 12h后,HO-1的mRNA表达水平较对照组明显升高(P值均＜ 0.01).槲皮素对肝细胞HO-1的诱导被PD98059抑制(0.79±0.05与0.16±0.02,t=19.520,P＜0.01),胞核内Nff2蛋白的表达也被PD98059明显抑制(0.14±0.04与0.04±0.01,t=4.114,P＜0.05).结论 槲皮素可能通过细胞外信号调节激酶/Nrf2信号转导通路诱导大鼠原代肝细胞HO-1的表达.“,”Objective To investigate the possible molecular mechanisms of heme oxygenase-1 (HO-1) induction by quercefin using rat primary hepatocyt.Methods Sprague-Dawley rat primary hepatocytes were isolated using a two-step collagenase perfusion technique and treated with quercetin at various doses (25-200μnol/L) and times (2-12 h).To investigate the roles of various signaling pathways,the hepamcytes were pretreated with 50 μmol/L quercetin plus an extracellular signal-regulated kinase (ERK) inhibitor (PD98059 at 10μmol/L),a p38 inhibitor (SB203580 at 10 μmol/L),a c-Jun N-terminal kinase inhibitor (SP600125 at 10 μmol/L)or a phosphatidylinositol 3-kinase inhibitor (Wortmannin at 1 rnol/L) for 12 h.Changes in the mRNA and protein levels of HO-1 and nuclear factor,ethryroid-2 related factor 2 (Nrf2) were detected by RT-PCR and western blotting.Results After 4-12 h of treatment with quercetin at all concentrations,the HO-1 mRNA level in hepatocytes had increased significantly (vs.untreated control cells; allP ＜ 0.01).The quercetin-induced HO-1expression and Nrf2 translocation into the nucleolus was inhibited by PD98059.Conclusion Quercetin may induce HO-1 expression via the ERK/Nrf2 signaling transduction pathway.