中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2013年
11期
865-868
,共4页
侯伟%刘爽%姚平%刘烈刚%覃华
侯偉%劉爽%姚平%劉烈剛%覃華
후위%류상%요평%류렬강%담화
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
肝細胞%有絲分裂素激活蛋白激酶類%血紅素氧化酶(脫環)%槲皮素%覈因子E2相關因子2
간세포%유사분렬소격활단백격매류%혈홍소양화매(탈배)%곡피소%핵인자E2상관인자2
Hepatocytes%Mitogen-activated protein kinases%Heme oxygenase(decyclizing)%Quercetin%NF-E2 related factor 2
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.
目的 研究槲皮素對大鼠原代肝細胞中Ⅰ型血紅素氧化酶(HO-1)誘導的分子機製.方法 二步膠原酶技術分離培養大鼠原代肝細胞.不同劑量槲皮素作用大鼠原代肝細胞不同時間後,用RT-PCR方法檢測HO-1的mRNA錶達;50μmol/L槲皮素分彆與10μmol/L PD98059、10μ mol/L SB203580、10 μ mol/L SP600125、1μmol/L Wormannin共孵育原代大鼠肝細胞12h後,用RT-PCR、Western blot法檢測HO-1 mRNA和覈因子E2相關因子2(Nrf2)蛋白錶達水平的變化.對數據進行單因素方差分析、Bonferroni t檢驗.結果 大鼠原代肝細胞經25 ～ 200μmol/L槲皮素處理12h,或用50μmol/L槲皮素處理4～ 12h後,HO-1的mRNA錶達水平較對照組明顯升高(P值均＜ 0.01).槲皮素對肝細胞HO-1的誘導被PD98059抑製(0.79±0.05與0.16±0.02,t=19.520,P＜0.01),胞覈內Nff2蛋白的錶達也被PD98059明顯抑製(0.14±0.04與0.04±0.01,t=4.114,P＜0.05).結論 槲皮素可能通過細胞外信號調節激酶/Nrf2信號轉導通路誘導大鼠原代肝細胞HO-1的錶達.
목적 연구곡피소대대서원대간세포중Ⅰ형혈홍소양화매(HO-1)유도적분자궤제.방법 이보효원매기술분리배양대서원대간세포.불동제량곡피소작용대서원대간세포불동시간후,용RT-PCR방법검측HO-1적mRNA표체;50μmol/L곡피소분별여10μmol/L PD98059、10μ mol/L SB203580、10 μ mol/L SP600125、1μmol/L Wormannin공부육원대대서간세포12h후,용RT-PCR、Western blot법검측HO-1 mRNA화핵인자E2상관인자2(Nrf2)단백표체수평적변화.대수거진행단인소방차분석、Bonferroni t검험.결과 대서원대간세포경25 ～ 200μmol/L곡피소처리12h,혹용50μmol/L곡피소처리4～ 12h후,HO-1적mRNA표체수평교대조조명현승고(P치균＜ 0.01).곡피소대간세포HO-1적유도피PD98059억제(0.79±0.05여0.16±0.02,t=19.520,P＜0.01),포핵내Nff2단백적표체야피PD98059명현억제(0.14±0.04여0.04±0.01,t=4.114,P＜0.05).결론 곡피소가능통과세포외신호조절격매/Nrf2신호전도통로유도대서원대간세포HO-1적표체.
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.
