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引用本文:郑 鹏, 王 波, 王 前.沙棘果油对过氧化氢诱导氧化损伤的保护作用[J].广西植物,2020,40(9):1349-1356.[点击复制]
ZHENG Peng, WANG Bo, WANG Qian.Protective effects of sea buckthorn fruit oil on oxidative damage induced by hydrogen peroxide[J].Guihaia,2020,40(9):1349-1356.[点击复制]
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沙棘果油对过氧化氢诱导氧化损伤的保护作用
郑 鹏1, 王 波2*, 王 前1
1. 西北濒危药材资源开发国家工程实验室/陕西师范大学生命科学学院, 西安 710119;2. 西安文理学院 生物与环境工程学院/西安市秦岭天然产物开发与抗癌类创新药物研究重点实验室, 西安 710065
摘要:
沙棘(Hippophae rhamnoides)是一种具有药用价值的植物,沙棘果油具有抗氧化、抗炎症及抗肿瘤等多种药理作用。为了探讨沙棘果油对H2O2造成氧化性损伤的细胞生长的影响及其抗氧化性,该研究选择H2O2对RAW264.7细胞氧化损伤模型,通过DPPH(1,1-二苯基-2-三硝基苯肼)自由基清除实验检测沙棘果油体外抗氧化能力,用[3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐]MTT法和流式细胞仪检测超氧化物阴离子荧光探针(DHE)信号,分别检测不同浓度沙棘果油对H2O2损伤细胞的存活率和超氧化物阴离子水平。结果表明:(1)在DPPH自由基清除实验中,当沙棘果油浓度小于4.9%时,沙棘果油的抗氧化能力大于维生素C。(2)通过MTT法发现,浓度为3.125%的沙棘果油对H2O2损伤细胞的存活率显著升高(P<0.01)。(3)从DHE检测发现,在同一检测时间点,随着沙棘果油浓度增加,DHE阳性细胞比例显著下降(P<0.01); 在不同检测时间点,随着沙棘果油浓度增加,DHE阳性细胞比例显著升高(P<0.01)。沙棘果油对过氧化氢诱导的RAW264.7细胞氧化损伤模型有一定修复作用,可能与细胞内超氧化物阴离子水平受到抑制有关,它具有抗氧化性损伤的潜能。
关键词:  沙棘果油, 抗氧化性, MTT法, DHE, 过氧化氢损伤模型
DOI:10.11931/guihaia.gxzw202004021
分类号:Q943.2
文章编号:1000-3142(2020)09-1349-08
基金项目:国家重点研发计划项目(2017YFC1701300); 中央高校基本科研业务费科技成果转化培育项目(GK201806005); 陕西省科技厅项目(2020JQ-888, 2019ZDLSF04-01-01, 2019SF-307); 西安市科技计划项目(CXY1531WL17)[Supported by the National Key R & D Program of China(2017YFC1701300); Central University Basic Research Service Fee Project(GK201806005); Key Special Project of Shaanxi Provincial Science and Technology Department(2020JQ-888, 2019ZDLSF04-01-01, 2019SF-307); Xi'an Science and Technology Plan Project(CXY1531WL17)]。
Protective effects of sea buckthorn fruit oil on oxidative damage induced by hydrogen peroxide
ZHENG Peng1, WANG Bo2*, WANG Qian1
1. National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest China/College of Life Sciences, Shaanxi Normal University, Xi'an 710119, China;2. College of Bioogical and Environmental Engineering/Key Laboratory of Natural Product Development and Anticancer Innovative Drug Research in Qinling, Xi'an University, Xi'an 710065, China 1. National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest China/College of Life Sciences, Shaanxi Normal University, Xi'an 710119, China; 2. College of Bioogical and Environmental Engineering/Key Laboratory of Natural Product Development and Anticancer Innovative Drug Research in Qinling, Xi'an University, Xi'an 710065, China
Abstract:
The sea buckthorn fruit(Hippophae rhamnoides)oil is medically claimed as having phytoantioxidant, antiinflammatory, and anticancerous properties in humans. To explore the hydrogen peroxide of the sea buckthorn fruit oil on mouse monocyte macrophages(RAW264.7 cells)oxidative damage model. By spectrophotometry, scavenging effects of sea buckthorn fruit oil on DPPH free radicals; In vitro antioxidant activity was assessed on RAW264.7 cells against hydrogen peroxide(H2O2)induced cytotoxicity. The antioxidant effect was determined by measuring the cell dye MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reducing assay. Then the survival rate of RAW264.7 cells was determined by MTT assay, and the expression of superoxide anion was determined by flow cytometry labeling with dihydroethidium(DHE). The results were as follows:(1)Compared with the antioxidant capacity of vitamin C, when the concentration of sea buckthorn fruit oil was less than 4.9%, the antioxidant capacity of sea buckthorn fruit oil was higher than vitamin C.(2)Comparing to the H2O2 group, 3.125% sea buckthorn fruit oil group had higher survival rate(P<0.01).(3)Comparing to the H2O2 group, the proportion of DHE-positive cell was significantly decreased in the same time(P<0.01)and it was negative effect with concentration of sea buckthorn fruit oil, the proportion of DHE-positive cell was significantly increased at the same concentration(P<0.01), and which was related positive effect with concentration of sea buckthorn fruit oil(P<0.01). These results support the protective effects of sea buckthorn fruit oil against hydrogen peroxide-induced toxicity in vitro with the underlying mechanisms of inhibiting oxidative stress. Taken together, these results clearly indicate that sea buckthorn fruit oil has significant potential as a natural antioxidant agent, which could be a potential source for the discovery of natrual antioxidant health product.
Key words:  sea buckthorn fruit oil, oxidation resistance, MTT, DHE, hydrogen peroxide damage model
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