Page 135 - 《广西植物》2020年第9期
P. 135
1 3 5 0 广 西 植 物 40 卷
Abstract: The sea buckthorn fruit ( Hippophae rhamnoides) oil is medically claimed as having phytoantioxidantꎬ
antiinflammatoryꎬ 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 spectrophotometryꎬ scavenging effects
of sea buckthorn fruit oil on DPPH free radicalsꎻ In vitro antioxidant activity was assessed on RAW264.7 cells against
hydrogen peroxide (H O ) induced cytotoxicity. The antioxidant effect was determined by measuring the cell dye MTT
2 2
[3 ̄(4ꎬ5 ̄dimethylthiazol ̄2 ̄yl) ̄2ꎬ5 ̄diphenyltetrazolium bromide] reducing assay. Then the survival rate of RAW264.7
cells was determined by MTT assayꎬ 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 Cꎬ
when the concentration of sea buckthorn fruit oil was less than 4.9%ꎬ the antioxidant capacity of sea buckthorn fruit oil
was higher than vitamin C. (2) Comparing to the H O groupꎬ 3.125% sea buckthorn fruit oil group had higher survival
2 2
rate(P<0.01). (3) Comparing to the H O groupꎬ the proportion of DHE ̄positive cell was significantly decreased in the
2 2
same time ( P < 0. 01) and it was negative effect with concentration of sea buckthorn fruit oilꎬ the proportion of
DHE ̄positive cell was significantly increased at the same concentration(P<0.01)ꎬ 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 togetherꎬ these results clearly indicate that sea buckthorn fruit oil has significant potential as a natural antioxidant
agentꎬ which could be a potential source for the discovery of natrual antioxidant health product.
Key words: sea buckthorn fruit oilꎬ oxidation resistanceꎬ MTTꎬ DHEꎬ hydrogen peroxide damage model
人体在代谢过程中会产生活性氧自由基(ROS)ꎬ 期研究发现ꎬ沙棘果油对酒精引起小鼠骨髓细胞
包括超氧阴离子(O ) 、羟自由基(OH)、过氧化氢 微核率和肝脏损伤有一定改善作用( 王波ꎬ2019)ꎮ
-
2
(H O ) 和脂质过氧化物的中间产物( Chauhan et 刘凤云(2005)研究发现ꎬ沙棘果油中含有棕榈油
2 2
al.ꎬ1997)ꎮ 在正常的生理情况下ꎬ线粒体需要消 酸、棕榈酸、油酸、亚油酸和不饱和脂肪酸ꎬ这些都
耗约 2% 的氧产生的 ROSꎬ由呼吸链底物端释放出 是人体不能自身合成的脂肪酸ꎬ此外还含有维生
-
电子还原氧分子单电子ꎬ产生 ROS 的前体 — O 素 C、维生素 E 和胡萝卜素等多种生物活性物质ꎮ
2
和 H O ꎮ 在很多疾病以及衰老时ꎬ体内氧自由基 魏安琪(2018)研究发现ꎬ黄酮类化合物、多酚类化
2 2
会产生异常积累ꎮ 过量的 ROS 会导致细胞损伤或 合物、维生素类、多肽类、多糖类物质等均可作为
细胞凋亡ꎮ ROS 的 活 泼 性 和 氧 化 反 应 能 力 都 很 天然抗氧化剂使用ꎮ 赵二劳等(2017) 研究发现ꎬ
强ꎬ能通过氧化作用攻击细胞的生物大分子物质( 沙棘果油在体外具有抗氧化性ꎬ但此研究仅利用
核酸、蛋白质、糖类、脂质) ꎮ 当人体处于亚健康 DPPH 自由基清除实验和 Fenton 实验在体外检测
状态时ꎬ自由基异常累积会引发疾病ꎮ 近年来ꎬ天 了沙棘果油的抗氧化活性ꎬ而其在体内的抗氧化
然抗氧化剂因其安全、高效、能满足亚健康人群日 效果仍不清楚ꎮ 然而ꎬ目前对日常可摄入食品中
常保健需求的特点而受到广泛关注ꎮ 的天然抗氧化成分含量的研究较少ꎬ尤其是在低
沙棘(Hippophae rhamnoides) 是一种具有药用 浓度下的抗氧化能力研究更为缺乏ꎬ沙棘果油作
价值的植物ꎬ为多年生落叶灌木或小乔木ꎬ适应性 为常用的保健品ꎬ有必要在细胞水平上对其抗氧
强ꎬ能显著改善生态环境( 丁健等ꎬ2016)ꎮ 沙棘果 化能力进行深入研究ꎮ
油是从沙棘的果肉中提取的油液ꎬ是沙棘的精华ꎬ ROS 具有性质活泼和反应性强的特点ꎬ既易
食用安全性高(林黎等ꎬ2018)ꎮ 目前ꎬ沙棘对调节 还原又易氧化ꎬ但其本身并不稳定ꎮ 对其检测的
血脂 ( 边 庆 荣 等ꎬ 1999)、 促 进 微 循 环 ( 吴 英 等ꎬ 手段有以下几种:电子自旋共振法( ESR)、高效液
2000)、抗炎、调节免疫(车锡平等ꎬ2000)以及抗癌 相色谱法、气相色谱法、化学发光法、荧光分析法、
(李忌等ꎬ2008) 等方面的研究已有报道ꎮ 我们前 分光光度法以及电化学方法ꎮ 但是ꎬ这些检测手