A comprehensive antioxidant evaluation was performed on enzymatic hydrolysates of Stichopusjaponicus (S. japonicus) using Vero cells and zebrafish models for in vitro and in vivo studies, respectively. S. japonicus was hydrolyzed with food-grade enzymes (alcalase, α-chymotrypsin, flavourzyme, kojizyme, neutrase, papain, pepsin, protamex, and trypsin), and the free radical scavenging activities were screened via electron spin resonance (ESR) spectroscopy. According to the results, the enzymatic hydrolysates contained high protein and relatively low polysaccharide and sulfate contents. Among these hydrolysates, the α-chymotrypsin assisted hydrolysate from S. japonicus (α-chy) showed high yield and protein content, and strong hydroxyl radical scavenging activity. Therefore, α-chy was chosen for further purification. The α-chy was fractionated by ultrafiltration into three ultrafiltration (UF) fractions based on their molecular weight: >10 kDa (α-chy-I), 5–10 kDa (α-chy-II), and <5 kDa (α-chy-III), and we evaluated their antioxidant properties in H₂O₂ exposed Vero cells. The α-chy and its UF fractions significantly decreased the intracellular reactive oxygen species (ROS) generation and increased cell viability in H₂O₂ exposed Vero cells. Among them, α-chy-III effectively declined the intracellular ROS levels and increased cell viability and exhibited protection against H₂O₂ induced apoptotic damage. Furthermore, α-chy-III remarkably attenuated the cell death, intracellular ROS and lipid peroxidation in H₂O₂ exposed zebrafish embryos. Altogether, our findings demonstrated that α-chy and its α-chy-III from S. japonicus possess strong antioxidant activities that could be utilized as a bioactive ingredient for functional food industries.

中文翻译：

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刺参酶解产物对过氧化氢诱导的氧化应激的潜在抗氧化性能

物上的酶水解产物进行一个全面的抗氧化剂的评价刺参刺参（S.刺参），使用Vero细胞和斑马鱼的模型，分别用于体外和体内研究。用食品级酶（alcalase，α-胰凝乳蛋白酶，风味酶，曲霉酶，中性酶，木瓜蛋白酶，胃蛋白酶，普他莫司和胰蛋白酶）水解日本血吸虫，然后通过电子自旋共振（ESR）光谱筛选自由基清除活性。根据结果​​，酶水解产物包含高蛋白质和相对低的多糖和硫酸盐含量。在这些水解产物中，日本血吸虫的α-胰凝乳蛋白酶辅助水解产物（α-chy）具有较高的产量和蛋白质含量，并具有很强的清除羟自由基的活性。因此，选择α-chy进行进一步纯化。根据分子量，通过超滤将α-chy分为三个超滤（UF）馏分：> 10 kDa（α-chy-I），5–10 kDa（α-chy-II）和<5 kDa（α） -chy-III），我们评估了它们在暴露于H ₂ O _2的Vero细胞中的抗氧化性能。α-chy及其超滤组分在暴露于H ₂ O _2的Vero细胞中显着减少了细胞内活性氧（ROS）的产生并提高了细胞活力。其中，α-chy-III有效地降低了细胞内ROS水平并提高了细胞活力，并表现出针对H _2的保护作用O ₂诱导细胞凋亡。此外，α-chy-III显着减弱了暴露于H ₂ O _2的斑马鱼胚胎中的细胞死亡，细胞内ROS和脂质过氧化作用。总的来说，我们的发现表明日本血吸虫的α-chy及其α-chy-III具有很强的抗氧化活性，可以用作功能性食品工业的生物活性成分。