| 槲皮素－AAPH氧化产物结构鉴定及其对 HepG2细胞氧化还原状态的影响 |
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| 引用本文： | 温斐婷,成向荣,陈侃俊,王菲,乐国伟,施用晖.槲皮素－AAPH氧化产物结构鉴定及其对 HepG2细胞氧化还原状态的影响[J].安徽农业科学,2016,44(17). |
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| 作者姓名： | 温斐婷 成向荣 陈侃俊 王菲 乐国伟 施用晖 |
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| 作者单位： | 江南大学食品学院,江苏无锡,214122;江南大学食品学院,江苏无锡,214122;江南大学食品学院,江苏无锡,214122;江南大学食品学院,江苏无锡,214122;江南大学食品学院,江苏无锡,214122;江南大学食品学院,江苏无锡,214122 |
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| 摘 要： | 目的]探究AAPH诱导的槲皮素氧化产物对Hep G2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Hep G2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Hep G2细胞出现氧化应激。
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| 关 键 词： | 槲皮素 氧化产 HepG2细胞 氧化应激 |
Structure Identification of the Quercetin Oxidation Products Induced by AAPH and Effects on the Redox State of HepG2 Cells |
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| 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. |
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| Keywords: | Quercetin Oxidation products HepG2 cells Oxidative stress |
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