CAJ | 학술논문

研究姜黄素对Ⅱ相酶谷胱甘肽转移酶(GST)及NADP(H)醌氧化还原酶(NQO)活性的影响及其诱导机制.用光谱法检测细胞GST酶和NQO酶的活性,以及还原型谷胱甘肽(GSH)的含量;利用蛋白印迹法检测核转录因子Nrf2在胞浆与胞核的分布;采用凝胶电泳迁移率分析法(EMSA)检测Nrf2与Ⅱ相酶基因抗氧化反应序列(ARE)结合活性.不同浓度的姜黄素(10～30 μmol·L-1)刺激结肠腺癌HT-29细胞后,能显著诱导GST酶及NQO酶活性的增加,同时能迅速提高细胞内GSH的含量;蛋白印迹和凝胶电泳迁移率结果显示,姜黄素诱导细胞核内转录因子Nrf2积聚,Nrf2-ARE的结合活性增加.姜黄素诱导的Ⅱ相酶GST酶及NQO酶活性增加与促进转录因子Nrf2由胞浆向胞核发生转位分布和增强Nrf2-ARE结合活性有关.
연구강황소대Ⅱ상매곡광감태전이매(GST)급NADP(H)곤양화환원매(NQO)활성적영향급기유도궤제.용광보법검측세포GST매화NQO매적활성,이급환원형곡광감태(GSH)적함량;이용단백인적법검측핵전록인자Nrf2재포장여포핵적분포;채용응효전영천이솔분석법(EMSA)검측Nrf2여Ⅱ상매기인항양화반응서렬(ARE)결합활성.불동농도적강황소(10～30 μmol·L-1)자격결장선암HT-29세포후,능현저유도GST매급NQO매활성적증가,동시능신속제고세포내GSH적함량;단백인적화응효전영천이솔결과현시,강황소유도세포핵내전록인자Nrf2적취,Nrf2-ARE적결합활성증가.강황소유도적Ⅱ상매GST매급NQO매활성증가여촉진전록인자Nrf2유포장향포핵발생전위분포화증강Nrf2-ARE결합활성유관.
This study is to investigate the effect of curcumin on the induction of glutathione Stransferases (GST) and NADP(H):quinone oxidoreductase (NQO) and explore their possible molecular mechanism. The activity of GST, NQO and cellular reduced glutathione (GSH) content were measured by spectrophotometrical methods. Cellular changes in the distribution of NF-E2 related factor 2 (Nrf2) were detected by Western blotting analysis. Nrf2-AREs (antioxidant-responsive elements) binding activity was examined by electrophoretic mobility shift assay (EMSA). Treatment of HT-29 human colon adenocarcinoma cells with curcumin dramatically induced the activity of GST and NQO at the range of 10-30 μmol·L-1. Curcumin exposure caused a significant increase in cellular GSH content rapidly as early as 3 h. Moreover, curcumin triggered the accumulation of Nrf2 in nucleus, and increased Nrf2 content in ARE complexes. These results demonstrated that induction of GST and NQO activity by curcumin may be mediated by translocation of transcription factor Nrf2 from cytoplasm to nuclear and increased binding activity of Nrf2-ARE complexes.