槲皮素诱导大鼠肝细胞I型血红素氧化酶表达的分子机制  

Potential molecular mechanisms of quercefin-induced heme oxygenase-1 in rat primary hepatocytes

作　　者：候伟[1] 刘爽[2] 姚平[3] 刘烈刚[3] 覃华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,武汉430030 [2]湖北省疾病预防控制中心卫生监测检验防护所 [3]华中科技大学同济医学院公共卫生学院营养与食品卫生学系

出　　处：《中华肝脏病杂志》2013年第11期865-868,共4页Chinese Journal of Hepatology

基　　金：国家自然科学基金青年基金（30500405）

摘　　要：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10Ixmol／LPD98059、10μmol／LSB203580、10μmol／LSP600125、1μmol／LWortmannin共孵育原代大鼠肝细胞12h后，用RT-PCR、Westernblot法检测HO-1mRNA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、Bonferronif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土0．05与0．16±0．02，t=19．520，P〈0．01），胞核内Nrf2蛋白的表达也被PD98059明显抑制（0．14士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 hepatocytes. Methods Sprague-Dawley rat primaly hepatocytes were isolated using a two-step collagenase perfusion technique and treated with quercetin at various doses （25 - 200 μmol/L） and times （2 - 12 h）. To investigate the roles of various signaling pathways, the hepatocytes were pre- treated with 50 μaol/L quercetin plus an extracellular signal-regulated kinase （ERK） inhibitor （PD98059 at 10 μmol/L）, a p38 inhibitor （SB203580 at 10 μnol/L）, a c-Jun N-terminal kinase inhibitor （SP600125 at 10 μmol/L） or a phosphatidylinositol 3-kinase inhibitor （Wortmannin at 1 pmol/L） for 12 h. Changes in the mRNA and protein levels of HO- 1 and nuclear factor, ethryroid-2 related factor 2 （Nrt＇2） 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 quereetin-induced HO-1 expression and Nrf2 translocation into the nucleolus was inhibited by PD98059. Conclusion Quercetin may induce HO-1 expression via the ERK/Nrt2 signaling transduction pathway.

关 键 词：肝细胞 有丝分裂素激活蛋白激酶类 血红素氧化酶(脱环) 槲皮素 核因子E2相关因子2 

分 类 号：R318.08[医药卫生—生物医学工程]