紫芜菁乙醇提物对Caco-2细胞氧化损伤的保护作用
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宋家乐(1983-),男,博士,副教授,研究方向:分子营养学和功能性食品学 通讯作者:周燕园(1976-),女,副教授,研究方向:食品理化检测新技术

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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


Protective Effect of Brassica rapa var L Ethanol Extract on Oxidative Damage of Caco-2 Cells
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    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(H2O2,150 μmol/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H2O2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H2O2引发的Caco-2细胞氧化应激损伤并降低炎症反应。

    Abstract:

    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.

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宋家乐,钱波,王程强,曾榛,桂中玉,周燕园.紫芜菁乙醇提物对Caco-2细胞氧化损伤的保护作用[J].现代食品科技,2018,34(6):32-38.

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  • 收稿日期:2018-01-22
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  • 在线发布日期: 2018-07-10
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