羊栖菜褐藻糖胶对AAPH诱导的斑马鱼氧化应激模型的保护作用

Protective Effects of Fucoidan Isolated from Sargassum fusiform on AAPH-induced Antioxidant Response in Zebrafish Model

目的:探究羊栖菜褐藻糖胶(SFPS)对2,2-偶氮二(2-甲基丙基咪)二盐酸盐(AAPH)诱导氧化应激斑马鱼模型的保护作用。方法:通过检测SFPS对2,2'-联氮-双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)及1,1-二苯基-2-苦基肼(DPPH)自由基的清除能力,来评价及验证SFPS的体外抗氧化能力,优化AAPH诱导斑马鱼氧化应激模型,并分别以三个表型指标胚胎存活率、卵黄囊大小和心跳速率以及斑马鱼体内细胞死亡率和活性氧(ROS)生成率来评价SFPS的体内抗氧化活性。结果:SFPS主要由75.30%±1.77%的总糖、21.39%±1.07%的硫酸根,1.78%±0.19%的蛋白质以及1.47%±0.02%的多酚组成,对DPPH、ABTS自由基清除的IC50值分别为0.59、0.69 mg/mL。此外,SFPS在50~200μg/mL范围内对斑马鱼胚胎无毒性,并对AAPH诱导引起的斑马鱼氧化损伤起到保护作用,且呈剂量依赖型。其中,在最优浓度200μg/mL显著提高斑马鱼胚胎存活率(100%,P<0.05),显著降低心跳速率(101.37%,P<0.05)和卵黄囊大小(111.80%,P<0.05),对斑马鱼体内细胞死亡和活性氧生成的抑制率最高分别可达70.55%和50.68%。结论:SFPS具有较强的体外抗氧化作用,并对AAPH诱导氧化损伤的斑马鱼表现出优越的体内抗氧化能力及氧化损伤修复能力,这表明SFPS作为天然抗氧化剂在保健食品及化妆品领域具有广泛应用前景。

Objective:A 2,2-azobis(2-methylpropylimidazole)dihydrochloride(AAPH)-stimulated zebrafish model was studied and used for investigating the antioxidant activities of a fucoidan(SFPS)isolated from Sargassum fusiform(S.fusiform).Methods:SFPS extracted from S.fusiform and the chemical composition of SFPS were determined.The scavenging capacities of SFPS on 1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt(ABTS)radicals were determined to evaluate its antioxidant activity in vitro.Then the AAPHinduced oxidative stress model of zebrafish was studied and optimized.Three phenotypic indexes(survival rate,yolk sac size and heartbeat rate)as well as levels of cell death and reactive oxygen species(ROS)were used to evaluate the antioxidant activity of SFPS in vivo.Results:The SFPS contained 75.30%±1.77%carbohydrates content,21.39%±1.07%sulfate content,1.78%±0.19%protein content and 1.47%±0.02%phenolic content.Compared with the crude polysaccharide(SFPSC),lower IC50 values of SFPS for DPPH and ABTS were determined to be 0.59 and 0.69 mg/mL,respectively.Besides,SFPS was nontoxic to zebrafish embryos in the range of 50~200μg/mL and dose-dependently protect zebrafishfrom AAPH induced oxidative damage.At the optimal concentration of 200μg/mL,the survival rate of zebrafish embryos was significantly increased(100%,P<0.05),the heartbeat rate and yolk sac size were significantly reduced(101.37%and 111.80%,respectively,P<0.05),and the inhibition rate of AAPH-induced cell death and ROS production in zebrafish were up to 70.55%and 50.68%,respectively.Conclusion:SFPS would have strong antioxidant activity in vitro,and had superior antioxidant ability and oxidative damage repair ability in vivo,thus indicated that SFPS as a natural antioxidant would have a wide application prospect in the field of health food and cosmetics.