仿刺参精、卵多肽体外抗氧化及对H2O2诱导巨噬细胞氧化损伤的保护作用

Antioxidative effects of Apostichopus japonicus sperm and egg polypeptide in vitro and their protective effects on H2O2-induced macrophage oxidative damage

中文关键词:  仿刺参精  仿刺参卵  多肽  抗氧化  巨噬细胞RAW 264.7  保护作用

英文关键词:Apostichopus japonicus sperm  Apostichopus japonicus egg  polypeptide  antioxidant  macrophage RAW 264.7  protective effects

中文摘要:

      目的 研究仿刺参精(Apostichopus japonicus sperm, AJS)和仿刺参卵(Apostichopus japonicus egg, AJE)多肽体外抗氧化及对H2O2诱导巨噬细胞氧化损伤的保护作用。方法 采用切向流超滤法对多肽进行分级制备, 通过测定清除1,1-二苯基-2-苦基苯肼自由基(1,1-diphenyl-2-picrylhydrazyl, DPPH·)、羟基自由基(hydroxyl radical, ·OH)和超氧阴离子自由基(O2-·)的能力来评价体外抗氧化活性, 并对多肽的相对分子质量(relative molecular mass, Mr)、氨基酸(amino acid, AA)组成及氧化稳定性进行检测分析, 通过构建氧化损伤细胞模型, 考察多肽对H2O2诱导RAW 264.7巨噬细胞氧化损伤的保护作用。结果 AJS和AJE多肽均具有较强的清除DPPH·、·OH和O2-·能力, 并且在一定范围内呈现量效关系, Mr<1×103 Da的多肽AJS1和AJE1抗氧化活性表现更强, 两者Mr 95.56%和91.89%在1×103 Da以内, 疏水性AA和酸性AA比例均较高, 并具有极佳的耐盐性和热稳定性, 但在贮藏和加工过程中仍要注意过酸环境和金属离子对氧化稳定性的影响。所有多肽组对巨噬细胞不存在毒性影响, 并且多肽质量浓度为200、400 μg/mL时均对H2O2损伤的RAW 264.7巨噬细胞体现出显著的保护作用(P<0.05)。结论 仿刺参精、卵多肽具有良好的体外抗氧化和对氧化损伤细胞的保护作用, 具有开发为功能性食品、医药和化妆品等的潜力。

英文摘要:

      Objective To study the antioxidative effects of Apostichopus japonicus sperm (AJS) and Apostichopus japonicus egg (AJE) polypeptide in vitro and their protective effects on H2O2-induced macrophage oxidative damage. Methods The polypeptides were prepared by tangential flow ultrafiltration, and their antioxidant activities in vitro were evaluated by scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH·), hydroxyl radical (·OH) and O2-·. Then the relative molecular mass (Mr), amino acid (AA) composition and oxidative stability of the polypeptide were detected and analyzed. By constructing an oxidative damage cell model, the protective effect of the polypeptide on H2O2-induced oxidative damage in RAW 264.7 macrophage was investigated. Results AJS and AJE polypeptides had strong scavenging abilities of DPPH·, ·OH and O2-·, and showed a dose-effect relationship within a certain range. The peptides AJS1 and AJE1 with Mr<1×103 Da showed stronger antioxidant activity, both Mr 95.56% and 91.89% were within 1×103 Da, and the proportion of hydrophobic AA and acidic AA were higher. They had excellent salt resistance and thermal stability, but the influence of peracid environment and metal ions on oxidative stability should be paid attention to during storage and processing. All peptide groups had no toxic effects on macrophages, thereafter the peptide mass concentrations of 200, 400 μg/mL showed significant protective effects on H2O2-injured RAW 264.7 macrophages (P<0.05). Conclusion AJS and AJE polypeptide has good in vitro antioxidant and protective effects on oxidatively damaged cells. Therefore they have the potential to be developed into functional foods, medicines and cosmetics.