基于秀丽隐杆线虫模型探讨白芥子水提物抗氧化作用及机制

Study on antioxidative effect and mechanism of water extracts of white mustard seed based on Caenorhabditis elegans model

中文关键词:  白芥子  抗氧化  作用机制  氧化应激

英文关键词:white mustard seed  antioxidant  mechanism  oxidative stress

中文摘要:

      目的 研究白芥子水提物抗氧化活性, 探讨其抗氧化的相关机制。方法 采用1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)自由基、羟基自由基、铁离子还原能力3种方法对其体外抗氧化活性进行评价。采用秀丽隐杆线虫野生型N2线虫观察白芥子水提物对线虫寿命、运动能力、生殖能力的影响; 通过氧化应激、热应激、脂褐素和活性氧(reactive oxygen species, ROS)评估其对线虫抗氧化能力的影响。采用实时荧光定量技术测定白芥子水提物作用下线虫抗氧化、胰岛素信号途径相关基因表达情况。结果 白芥子水提物具有一定的DPPH自由基、羟基自由基清除能力。通过氧化应激筛选出最佳给药质量浓度为10 mg/mL, 且该给药质量浓度对线虫的运动能力和生殖能力无影响(P>0.05), 并能够显著延长线虫的平均寿命和中位寿命(P<0.01); 10 mg/mL给药组显著提高线虫抗热应激能力, 降低脂褐素积累和ROS水平; 同时上调daf-16、sod-3、clt-2、skn-1的mRNA表达量, 下调daf-2的mRNA的表达量。结论 白芥子具有良好的体内外抗氧化能力, 可能与调控胰岛素/胰岛素样生长因子(insulinlike growth factor, IGF)-1信号(insulin/IGF-1 signaling, IIS)通路上的基因表达有关。

英文摘要:

      Objective To study the antioxidant activity of the water extract of white mustard seed and explore its antioxidant mechanism. Methods The antioxidant activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydroxyl radical and iron ion reducing ability in vitro was evaluated. The wild-type N2 nematode of Caenorhabditis elegans was used to observe the effect of the water extracts of white mustard seed on the life span, locomotor ability and reproductive ability of the nematode. The effects on the antioxidant capacity of nematodes were assessed by oxidative stress, heat stress, lipofuscin and reactive oxygen species (ROS). The expression levels of genes related to antioxidant and insulin signaling pathways in Caenorhabditis elegans induced by the water extracts of white mustard seed were determined by real-time fluorescence quantitative analysis. Results The water extracts of white mustard seed had certain DPPH free radical and hydroxyl radical scavenging activities. The optimal concentration of 10 mg/mL was screened out by oxidative stress and it had no effect on the locomotor ability and reproductive ability of Caenorhabditis elegans (P>0.05) and could significantly prolong the average and median life span of Caenorhabditis elegans (P<0.01). The 10 mg/mL treatment group significantly improved the heat stress resistance of Caenorhabditis elegans and reduced the accumulation of lipofuscin and ROS level. At the same time, the mRNA expression levels of daf-16, sod-3, clt-2 and skn-1 were increased, and the mRNA expression level of daf-2 was decreased. Conclusion Mustard seed has good antioxidant capacity in vitro and in vivo, which may be related to the expression of genes regulating insulin like growth factor (IGF)-1 signaling (IIS) pathway.