Nutrition and Food Technology Group, Faculty of Pharmaceutical and Food Sciences, University of Antioquia, Medellin 050010, Colombia.
Nutrition and Food Science Area, Faculty of Pharmacy and Food Sciences, University of Valencia, Avda. Vicente Andrés Estellés s/n, 46100 Burjassot, Valencia, Spain.
Nutrients. 2024 Oct 27;16(21):3654. doi: 10.3390/nu16213654.
BACKGROUND/OBJECTIVES: When prooxidants outweigh antioxidants, oxidative stress can occur, causing an accumulation of reactive oxygen species (ROS). This process can lead to cellular damage and plays a role in the development of numerous health conditions. This study aimed to investigate the cytoprotective effects on differentiated Caco-2 cells of hydrolysates derived from the red Californian worm (WH) and their fractions, identify the peptides responsible for this effect, and elucidate the mechanisms involved.
The WH was obtained through hydrolysis with Alcalase 2.4 L and subsequently fractionated to two fractions (F > 3 kDa and F < 3 kDa) using a ceramic membrane with a molecular weight cutoff of 3 kDa. The peptides found in the F < 3 kDa fraction, demonstrating the highest cytoprotective activity, were then sequenced via liquid chromatography-mass spectrometry analysis (LC-MS/MS), and molecular docking was conducted to elucidate the underlying antioxidant mechanisms.
The hydrolysate of and its F < 3 kDa fraction exhibited no cytotoxicity, protected the cells from HO-induced oxidative stress (50% increase viability), preserved cell viability by restoring their redox status (ROS: 20% decrease, and glutathione (GSH): recovered to basal control levels) and cell cycle distribution, and decreased apoptosis (16%). Twenty-eight peptides were identified, with five showing antioxidant activity through stable interactions with myeloperoxidase (MPO) and Kelch-like ECH-associated protein 1 (Keap-1), KPEDWDDR being the peptide that presented the highest affinity with both molecules (-7.9 and -8.8 kCal/mol, respectively).
These results highlight the WH as a potential source of bioactive peptides for the management of oxidative stress.
背景/目的：当促氧化剂超过抗氧化剂时，可能会发生氧化应激，导致活性氧（ROS）的积累。这个过程会导致细胞损伤，并在许多健康状况的发展中发挥作用。本研究旨在研究来源于加利福尼亚红虫（WH）及其水解产物和级分的对分化的 Caco-2 细胞的细胞保护作用，鉴定负责这种作用的肽，并阐明所涉及的机制。
通过 Alcalase 2.4 L 水解获得 WH，然后使用分子量截止值为 3 kDa 的陶瓷膜将其分为两个级分（F > 3 kDa 和 F < 3 kDa）。在 F < 3 kDa 级分中发现的具有最高细胞保护活性的肽，然后通过液相色谱-质谱分析（LC-MS/MS）进行测序，并进行分子对接以阐明潜在的抗氧化机制。
和其 F < 3 kDa 级分的水解产物没有细胞毒性，可保护细胞免受 HO 诱导的氧化应激（活力增加 50%），通过恢复其氧化还原状态（ROS：减少 20%，谷胱甘肽（GSH）：恢复到基础对照水平）和细胞周期分布来维持细胞活力，并减少凋亡（16%）。鉴定出 28 个肽，其中 5 个通过与髓过氧化物酶（MPO）和 Kelch 样 ECH 相关蛋白 1（Keap-1）的稳定相互作用显示出抗氧化活性，KPEDWDDR 是与这两种分子结合亲和力最高的肽（分别为-7.9 和-8.8 kCal/mol）。
这些结果强调了 WH 作为管理氧化应激的生物活性肽的潜在来源。
