槲皮素对小鼠体内和体外NIH-3T3细胞的抗氧化作用及其机理  

Effect and underlying mechanism of antioxidant quercetin in mouse and in NIH-3T3 cells

作　　者：宫璀璀[1] 郑乃刚[2,3] 吴景兰[2] 何培霞[1] 王一菱[2] 

机构地区：[1]中国人民解放军第153医院检验中心,河南郑州450042 [2]郑州大学分子细胞生物学研究中心,河南郑州450052 [3]郑州大学基础医学院肿瘤基础学教研室,河南郑州450001

出　　处：《基础医学与临床》2009年第1期52-58,共7页Basic and Clinical Medicine

基　　金：河南省基础与前沿技术研究计划项目(072300410440)

摘　　要：目的比较槲皮素对小鼠体内和体外H2O2处理前/后的NIH-3T3细胞抗氧化效应的差异并探讨其作用机理。方法将NIH-3T3细胞分为4组,槲皮素前保护组(Qb)、槲皮素后保护组(Qa)、H2O2组(H2O2)和对照组(C)。用试剂盒检测细胞内T-AOC、SOD、GSH-Px、GSH、MDA、NOS和NO2-/NO3-的水平;MTT和TUNEL法检测细胞凋亡率和生存率;免疫印迹和免疫组化技术检测细胞的cyclin D1、PTEN、NF-κB、HSP-70、BCl-2、BAX、及caspase-3的表达。另将20只Wistar大鼠等分为对照组和实验组,检测槲皮素灌胃前、后1、2及24 h血浆内T-AOC、SOD、GSH-Px、GSH、MDA、NOS和NO2-/NO3-的水平。结果H2O2组细胞凋亡率增高,cyclin D1、PTEN及BCl-2的表达下调;BAX、HSP-70、NF-κB及caspase-3的表达上调;T-AOC,SOD,GSH-Px及GSH水平降低,NOS、NO2-/NO3-及MDA增高。动物灌胃后1 h、2 h血浆中T-AOC、SOD、GSH-Px、GSH,NOS及NO2-/NO3-的水平增高,MDA降低;24 h后基本恢复。结论槲皮素在体内外皆可通过调节抗氧化酶体系发挥抗氧化作用,其效应依据H2O2处理与否和处理前/后等细胞的异质性而呈现一定差异。Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment, to explore the underlying mechanism for the quercetin antioxidant Methods The cultured NIH-3T3 cells were randomly divided into 4 groups： quercetin （Q） pre-protective group （Qb） firstly treated with quereetin for 24 h followed by incubation with H2O2 for 30 min; post- protective group （ Qa ） treated with H2O2 for 30 rain followed by incubation with quercetin for 24 h ; H2O2 group （ H2O2 ） after exposure to H2O2 for 30 min, incubated with DMEM medium and the control group （C） only cultured with DMEM medium. The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell suspension samples. The expression of cyclin D1, PTEN, NF-κB, HSP-70, BCl-2, BAX and caspase-3 were examined with immunocytochemistry and immunoblotting. Besides, 20 Wistar rats were divided into control group and experimental group, the latter was given with quercetin in the doze of 0. 13 mmol/kg. The levels of T-AOC ,SOD, GSH-Px,GSH,MDA, NOS and NO2^-/NO3^- were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h, 2 h and after 24 h. Results When the Qb group was compared with H2O2 or Qa group, the survival rate was higher and the apoptotic rate was lower. When the H2O2 group was compared with C group,the expression of cyclin D1 ,PTEN or BCl-2 was down-regulated ;while that of BAX,HSP-70 ,NF-κB or caspase-3 was up-regulated; the level of T-AOC, SOD,GSH-Px or GSH was decreased; that of NOS,NO2^-/NO3^- or MDA enhanced in the cleft NIH-3T3 cells. When the plasma level of the anti-oxidative enzyme system prior to- compared with post-lh and 2h-treatment with Q, the level of T-AOC, SOD, GSH-Px and GSH, especially the former two, were higher; MDA, lower; NOS or NO2^-/NO3^- promoted. However, the above parameters basically became normal 24 h after treatment with Q. Conclusion Quer

关 键 词：槲皮素 H2O2氧化应激 抗凋亡信号传导 NIH-3T3细胞系 抗氧化酶体系 

分 类 号：R285.5[医药卫生—中药学]