碱性蛋白酶水解大豆水解物的抗氧化活性及对人肠道上皮 Caco-2 细胞的保护作用。

Antioxidant activity and protective effects of Alcalase-hydrolyzed soybean hydrolysate in human intestinal epithelial Caco-2 cells.

机构信息

College of Food Science, Northeast Agricultural University, China; National Research Center of Soybean Engineering and Technology, Harbin, 150030, China.

出版信息

Food Res Int. 2018 Sep;111:256-264. doi: 10.1016/j.foodres.2018.05.046. Epub 2018 May 21.

DOI:10.1016/j.foodres.2018.05.046

PMID:30007684

Abstract

Soybeans are known as a promising source of bioactive peptides. However, knowledge on the antioxidant behaviors of soybean protein hydrolysate (SPH) in the human intestinal epithelium is limited. In this study, SPH was prepared with Alcalase and subsequently ultrafiltered into four peptide fractions as SPH-I (<3 kDa), SPH-II (3-5 kDa), SPH-III (5-10 kDa) and SPH-IV (>10 kDa). The antioxidant properties of SPH and membrane fractions were investigated using different chemical assays and their protective effects against oxidative stress were evaluated using HO-stressed human intestinal Caco-2 cells. Results showed that SPH-I exhibited the strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (IC = 2.56 mg/mL) and reducing capacity while SPH-III had the best metal ion-chelating activity (IC = 0.29 mg/mL). Both SPH and the peptide fractions dose-dependently suppressed intracellular reactive oxygen species (ROS) accumulation induced by HO in Caco-2 cells, but the strongest inhibitory effect was observed for SPH-I. Amino acid (AA) results revealed that SPH-I was rich in hydrophobic and antioxidant AAs, which could contribute to its stronger antioxidant properties. Additionally, SPH-I protected Caco-2 cells from HO-induced oxidative stress via inhibiting lipid peroxidation and stimulating antioxidant enzyme activities. These results suggest that SPH-I and constitutive peptides can be beneficial ingredients with antioxidant properties and protective effects against ROS-mediated intestinal injury.

摘要

大豆被认为是生物活性肽的有前途的来源。然而，关于大豆蛋白水解物（SPH）在人体肠道上皮中的抗氧化行为的知识有限。在这项研究中，使用 Alcalase 制备 SPH，并随后通过超滤将其分为四个肽级分，即 SPH-I（<3 kDa）、SPH-II（3-5 kDa）、SPH-III（5-10 kDa）和 SPH-IV（>10 kDa）。使用不同的化学测定法研究了 SPH 和膜级分的抗氧化特性，并使用 HO 应激的人肠道 Caco-2 细胞评估了它们对氧化应激的保护作用。结果表明，SPH-I 表现出最强的 2,2-二苯基-1-苦基肼（DPPH）自由基清除活性（IC = 2.56 mg/mL）和还原能力，而 SPH-III 具有最强的金属离子螯合活性（IC = 0.29 mg/mL）。SPH 和肽级分均以剂量依赖性方式抑制 HO 在 Caco-2 细胞中诱导的细胞内活性氧（ROS）积累，但 SPH-I 的抑制作用最强。氨基酸（AA）结果表明，SPH-I 富含疏水性和抗氧化性氨基酸，这可能有助于其更强的抗氧化特性。此外，SPH-I 通过抑制脂质过氧化和刺激抗氧化酶活性来保护 Caco-2 细胞免受 HO 诱导的氧化应激。这些结果表明，SPH-I 和组成肽可以是具有抗氧化特性和保护作用的有益成分，可防止 ROS 介导的肠道损伤。