枸杞对秀丽隐杆线虫寿命和产卵的影响及其抗氧化作用

目的:研究枸杞对秀丽隐杆线虫(以下简称线虫)寿命和产卵的影响,探讨其抗氧化的相关机制。方法:采用铁离子还原/抗氧化能力法体外检测,比较了宁夏中宁、青海、甘肃3个产地的枸杞抗氧化能力的差异。采用活性氧(reactive oxygen species,ROS)-敏感染料2’,7’-二氯-二氢荧光素乙酰乙酸盐检测枸杞对线虫体内ROS的影响,同时采用实时荧光定量聚合酶链式反应检测枸杞对线虫体内相关抗氧化酶水平的影响。将同步化后L4时期的线虫分别放置于对照或添加枸杞提取物的NGM平板上培养,检测枸杞对线虫产卵和寿命的影响。采用两种转基因(Pmyo3::Hy Per和ro GFP2::Orp1)线虫为模型检测枸杞提取物对线虫胞质中H_2O_2的清除能力,采用荧光染料Mito Sox Red检测枸杞提取物对线虫细胞线粒体中超氧阴离子自由基的清除能力。结果:3个产地的枸杞均具有很强的体外抗氧化能力,它们之间没有明显差异。枸杞可以降低线虫体内ROS水平以及提高抗氧化酶的水平。枸杞对线虫寿命和产卵能力并没有明显的影响。枸杞可以清除线粒体超氧阴离子自由基,但不影响胞质H_2O_2水平。结论:枸杞在体外与体内均有一定的抗氧化能力,其主要通过清除线粒体超氧阴离子自由基在体内发挥抗氧化作用,枸杞对线虫的寿命和产卵数没有明显影响。

Effect of Goji Berries (Lycium barbarum) on Lifespan and Spawning of Caenorhabditis elegans and Its Antioxidant Capacity

Objective: To evaluate the effect of Goji berries (Lycium barbarum) on the lifespan and spawning of Caenorhabditis elegans, and to explore its antioxidant capacity. Methods: The ferric reducing/antioxidant power (FRAP) method was used to assess and compare the antioxidant capacity of Goji berries from three different areas of Ningxia, Qinghai and Gansu provinces. The effect of Goji berries on the level of reactive oxygen species (ROS) in nematodes was detected using the fluorescent dye 2’,7’-dichlorofluorescein diacetate, and the effect on the expression of antioxidant enzymes in nematodes was evaluated by real-time quantitative polymerase chain reaction. In addition, in order to determine the effect of Goji berries on the lifespan and spawning, L4 stage nematodes were placed on NGM plates and incubated with and without added Goji berries. The scavenging capacity of Goji berries against H2O2 in the cytoplasm was detected using two transgenic nematodes: Pmyo3::HyPer and roGFP2::Orp1. The fluorescent dye MitoSox Red was used to measure the scavenging capacity against superoxide anion radical in the mitochondria. Results: Goji berries from all three areas had strong antioxidant activity in vitro with no significant difference being found between them. Goji berries could reduce the level of ROS and increase the levels of antioxidant enzymes in nematodes. However, Goji berries had no significant effect on the lifespan or spawning ability. Goji berries could scavenge mitochondrial superoxide anion radicals but not cytoplasmic H2O2. Conclusion: Goji berries can exert antioxidant capacity in vitro and in vivo, and its antioxidant effect in vivo is achieved by reducing the level of mitochondrial superoxide anion radicals. In addition, Goji berries do not dramatically affect the lifespan or the total number of eggs laid in nematodes.