为进一步实现琼胶寡糖的高值化利用,探究琼胶寡糖(Agaro-oligosaccharides,AOS)对2,2-偶氮二(2-甲基丙基咪)二盐酸盐(AAPH)诱导损伤后斑马鱼的抗氧化作用及机制。采用高效液相色谱法对酸法制备的琼胶寡糖组分进行分析。优化建立AAPH诱导的斑马鱼模型,并以斑马鱼胚胎存活率、活性氧生成率和细胞死亡率评价不同浓度琼胶寡糖的抗氧化活性。结果表明,琼胶寡糖主要由琼二糖(51.60%)、琼四糖(28.40%)和琼六糖(14.50%)组成,对斑马鱼胚胎添加琼胶寡糖浓度低于200 μg/mL时无致畸、致死毒性;琼胶寡糖(25、50、100 μg/mL)对最优AAPH诱导浓度(15 mmol/L)引起的氧化应激损伤斑马鱼模型有保护作用,且呈剂量依赖性。其中最优浓度100 μg/mL 琼胶寡糖能显著提高斑马鱼胚胎存活率(96.88%,p<0.05),显著降低AAPH引起的斑马鱼体内活性氧生成率(100.53%,p<0.05)和细胞死亡率(101.69%,p<0.05)。本研究通过斑马鱼模型首次揭示了琼胶寡糖可通过清除体内大量活性氧生成和阻止细胞死亡损伤的抗氧化机制实现提高其体内抗氧化活性,为其作为保健功能食品等应用提供了理论基础。
In order to further realize the high value utilization of agaro-oligosaccharides (AOS), antioxidant activities and mechanisms of AOS, 2, 2-azobis (2-methylpropylimidazole) dihydrochloride (AAPH) -induced zebrafish model were studied. The composition of AOS was determined by HPLC. The optimal AAPH induced zebrafish oxidative stress model was established, and the antioxidant activities of AOS with different concentrations were investigated by determination of embryo survival rate, reactive oxygen production rate, and cell death rate. The results showed that AOS was mainly composed of agarobiose (51.60%), agarotetraose (28.40%) and agarohexaose (14.50%). Zebrafish embryos with AOS concentration below 200 μg/mL had no teratogenic and lethal toxicity. AOS (25, 50, 100 μg/mL) had a protective effect on oxidative stress induced by AAPH (15 mmol/L) in a dose-dependent manner. The optimal concentration of 100 μg/mL AOS could significantly increase the survival rate of zebrafish embryos (96.88%, p<0.05), and could significantly reduce the reactive oxygen species production rate (100.53%, p<0.05) and cell death rate (101.69%, p<0.05) of AAPH-induced zebrafish.In this study, the zebrafish model was used to reveal for the first time. AOS had antioxidant activity in vivo by eliminating a large number of reactive oxygen species (ROS) production and preventing cell death and injury, which provided a theoretical basis for the application of AOS as a health food.