摘要
[目的]探究AAPH诱导的槲皮素氧化产物对HepG2细胞氧化还原状态的影响。[方法]用紫外－可见波长扫描、HPLC－MS方法分析AAPH和槲皮素在37℃下反应12 h生成的产物结构；以槲皮素－AAPH反应产物处理HepG2细胞24 h，测定其对细胞存活率、ROS水平及细胞总抗氧化能力（T－AOC）、谷胱甘肽过氧化物酶（GSH－Px）活性、超氧化物歧化酶（SOD）活性、GSH／GSSG比值及丙二醛（MDA）含量的影响，并测定Nrf2、Keap1、NQO1、Prdx1相关抗氧化基因的mRNA表达。[结果] AAPH与槲皮素体系在反应12 h后于294 nm波长处有特征吸收，LC－MS解析结果表明该体系反应生成了含4种主要氧化产物的混合物。与空白组相比，槲皮素与AAPH氧化产物孵育显著降低了细胞存活率，同时细胞内ROS水平和MDA含量升高，细胞内T－AOC水平、GSH－Px及SOD活性显著降低，并显著下调Nrf2、NQO1、Prdx1 mRNA表达，上调Keap1 mRNA表达。[结论]槲皮素氧化产物可造成HepG2细胞出现氧化应激。
Abstract
Objective] The aim was to explore how the quercetin oxidation products induced by AAPH influence the redox state of HepG2 cells.[ Method] Quercetin was mixed with APPH as oxidant under 37 ℃ for 12 h.The structural characterization were analyzed by UV scan and LC-MS.HepG2 cells were treated with reaction product of quercetin-AAPH and incubated for 24 h.Viable cells were harvested, and the cell viability, ROS level, glutathione ( GSH) and glutathione disulphide ( GSSG) content, malandialdehyde ( MDA) level, total antioxidant capacity ( T-AOC) , superoxide dismutase ( SOD) and catalase ( CAT) activity were assayed with the appropriate test kits, the oxidant stress related genes were assayed by DNA microarray.[Result] The chromatogram showed peaks eluting at 294 nm after 12 h reaction and the struc-tural analysis result indicated that mixture of four oxidation products were obtained in the reaction system.Compared with the blank group, Quercetin-AAPH oxidation products elevated the ROS and MDA generation, and reduced cell viability, T-AOC, CAT, GSH-Px and SOD and GSH/GSSG activity significantly.Nrf2, NQO-1, Prdx expression level was down-regulated and Keap-1, Nox-2, Cox-2 expression level was up-regulated in the meanwhile.[Conclusion] These findings indicate that quercetin oxidation products could prompt HepG2 cells to generate more oxidative stress.
国家科技期刊平台
NSTL
维普
万方数据