摘要
探讨紫芜菁乙醇提取物(BREE)的体外抗氧化能力及其对人肠上皮Caco-2细胞氧化损伤保护效果。二苯代苦味酰基自由基(DPPH)、羟基自由基(·OH)清除率与总还原力评估BREE的体外抗氧化力。过氧化氢(H_2O_2,150μmol/L)建立Caco-2细胞氧化损伤模型评估BREE的细胞保护效果。受损细胞生存率的影响以MTT法测定。细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化酶(GSH-Px)水平依说明用试剂盒测定。硫代巴比妥酸法测定丙二醛(MDA)含量。2’,7’-二氢二氯荧光黄双乙酸钠法测定细胞内活性氧(ROS)水平。酶联法测定白介素(Interlukin,IL)-1β和IL-8的水平。BREE具有较强的自由基(DPPH和·OH)清除力和总还原力。BREE能显著抑制H_2O_2造成的细胞死亡,提高受损细胞中抗氧化物酶(SOD、CAT和GSH-Px)活性,降低受损细胞内MDA与ROS的生成,并抑制IL-1β和IL-8的分泌。结果提示,BREE具有较强的体外抗氧化能力,能增强细胞内源性抗氧化酶的活力显著改善H_2O_2引发的Caco-2细胞氧化应激损伤并降低炎症反应。
The in vitro antioxidant activity and protective effect of Brassica rapa ethanol extracts（BREE） on oxidative damage of Caco-2 cells were investigated. In vitro antioxidant capability of BCEE was evaluated according to the 1,1-diphenyl-2-picrylhydrazyl（DPPH） and hydroxyl radicals（·OH） scavenging ability, and reducing power assay, respectively. Cellular protective effect of BREE was investigated using a model of intestinal epithelial Caco-2 cells induced oxidative stress by hydrogen peroxide（H2O2, 150 μmol/L） of. Cell viability was determined by MTT assay. The intracellular levels of superoxide dismutase（SOD）, catalase（CAT） and glutathione peroxidase（GSH-Px） were determined by commercial assay kits according the manuscripts. The intracellular levels of malondiadehycle（MDA） and reactive oxygen specisis（ROS） were determined by thiobarbituric acid reactive substance（TBARS） and dichloro-dihydro-fluorescein diacetate（DCFH-DA） assay, respectively. The levels of interlukin（IL）-1β and IL-8 were determined by enzyme linked immunosorbent assay（ELISA） kits. BREE exhibited a great activity of scavenging DPPH and ·OH radicals, and also showed a strong reducing power. BREE significantly increased the cell viability and activities of SOD, CAT and GSH-Px in cells. In addition, BREE also effectively decreased the intracellular levels of ROS and MDA, and reduced the secretion of IL-1β and IL-8 in Caco-2 cells treated with H2O2. These results suggested that the BREE exhibited a good in vitro antioxidant activity. BREE treatment could enhance the activities of endogenous antioxidant enzymes, which could attenuate the oxidative damage and reduce the inflammation reaction in Caco-2 cells treated with H2O2.
作者
宋家乐
钱波
王程强
曾榛
桂中玉
周燕园
SONG Jia-le;QIAN Bo;WANG Cheng-qiang;ZENG Zhen;GUI Zhong-yu;ZHOU Yan-yuan(School of Public and Health, Guilin Medical University, Guilin 541100, China;School of Public and Health, Southeast University, Nanjing 2 10009, China;Guangxi Colleges and University Key Laboratory of Preventive Medicine, Guilin Medical University, Guilin 541100, China;School of Pharmacy, Guilin Medical University, Guilin 541100, China)
出处
《现代食品科技》
EI
CAS
北大核心
2018年第6期32-38,共7页
Modern Food Science and Technology
基金
国家自然科学基金资助项目(81560530
81760589)
广西壮族自治区自然科学基金资助项目(2016GXNSFCA380026)
桂林医学院引进人才科研启动基金(04010150001)
2017年教育部中西部高等学校青年骨干教师国内访问学者计划资助
关键词
紫芜菁乙醇提取物
抗氧化能力
氧化应激
肠上皮细胞
炎症
brassica rapa var L ethanol extracts
antioxidant activity
oxidative stress
intestinal epithelial cells
inflammation
