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Mar Drugs
2019 Nov 14;1711:. doi: 10.3390/md17110642.
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Preparation of Antioxidant Peptide by Microwave- Assisted Hydrolysis of Collagen and Its Protective Effect Against H2O2-Induced Damage of RAW264.7 Cells.
Li Y
,
Li J
,
Lin SJ
,
Yang ZS
,
Jin HX
.
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Antioxidant peptides have elicited interest for the versatility of their use in the food and pharmaceutical industry. In the current study, antioxidant peptides were prepared by microwave-assisted alkaline protease hydrolysis of collagen from sea cucumber (Acaudina molpadioides). The results showed that microwave irradiation significantly improved the degree of hydrolysis of collagen and the hydroxyl radical (OH⋅) scavenging activity of hydrolysate. The content and OH⋅ scavenging activity of collagen peptides with molecular weight ≤ 1 kDa (CPS) in the hydrolysate obtained at 250 W increased significantly compared with the non-microwave-assisted control. CPS could scavenge OH⋅ and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical in a dose-dependent manner. The scavenging activity of OH⋅ and DPPH radical was 93.1% and 41.2%, respectively, at CPS concentration of 1 mg/mL. CPS could significantly promote RAW264.7 cell proliferation and reduce the Reactive Oxygen Species (ROS) level of H2O2-induced damage in RAW264.7 cells in a dose-dependent manner. Furthermore, all CPS-treated groups exhibited an increase in superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and a decrease in malondialdehyde (MDA) level compared with the control. These results showed that CPS could effectively protect RAW264.7 cells from H2O2-induced damage, implying the potential utilization of CPS as a natural antioxidant for food and pharmaceutical applications.
Figure 1. Effect of microwave power on OH· scavenging activity of collagen hydrolysate. The concentration of hydrolyzed product was 2 mg/mL. Values with different letters are significantly different (p < 0.05).
Figure 2. The OH· scavenging activity of peptides with different Mw (CPL, Mw > 5 kDa; CPM, 1 kDa < Mw ≤ 5 kDa; CPS, Mw ≤ 1 kDa) from hydrolysate of ASC-Am. (a–b) Values with different letters indicated significant differences in the same samples at different microwave powers (P < 0.05). (A–C) Values with different letters indicated significant differences in different samples at the same microwave powers (p < 0.05).
Figure 3. The scavenging activity of OH· (A) and DPPH (B) of CPS obtained by alkaline protease hydrolysis of ASC-Am at 250 W of microwave power. Ascorbic acid (AA) was used as a positive control. (a–b) Values with different letters indicated significant differences in different samples at the same concentrations (p < 0.05). (A–F) Values with different letters indicated significant differences in the same samples at the different concentrations (p < 0.05).
Figure 4. The effect of H2O2 on the viability of RAW264.7 cells. (a–g). Values with different letters indicated significant differences in the same time at different concentrations of H2O2 (p < 0.05). (A–C) Values with different letters indicated significant differences in the same concentrations of H2O2 at different times (p < 0.05).
Figure 5. Effect of CPS on viability of RAW264.7 cells. Values with different letters are significantly different (p < 0.05).
Figure 6. Effect of CPS on ROS levels in RAW264.7 cells induced by H2O2. (A) Control group; (B) model group (500 µM H2O2); (C) low-dose group (100 µg/mL CPS + 500 µM H2O2); (D) middle-dose group (150 µg/mL CPS + 500 µM H2O2); (E) high-dose group (200 µg/mL CPS + 500 µM H2O2).
Figure 7. Effect of CPS on levels of MDA (A), GSH-Px (B), and SOD (C) in RAW264.7 cells induced by H2O2. Model group (500 µM H2O2), low-dose group (100 µg/mL CPS + 500 µM H2O2), middle-dose group (150 µg/mL CPS + 500 µM H2O2), high-dose group (200 µg/mL CPS + 500 µM H2O2). Values with different letters are significantly different (p < 0.05).
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