荞麦多肽对RAW 264.7细胞氧化损伤的保护作用

Protective Effect of Buckwheat Antioxidant Peptides on Oxidative Damage of RAW 264.7 Macrophages

为提高荞麦蛋白生物利用度,利用Sephadex G-25凝胶层析和反向高效液相色谱(RP-HPLC)分离制备荞麦抗氧化多肽(BAPs),对其结构进行表征。通过H_(2)O_(2)诱导RAW 264.7巨噬细胞建立氧化应激模型,评价BAPs对细胞氧化损伤的保护作用。结果表明:经分离纯化得到的BAPs纯度达96.25%,其分子质量为5.8 ku。BAPs对DPPH·、ABTS+·、·OH清除能力及BAPs的还原力以VC当量抗氧化能力(VCEAC)分别表示为(12.12±0.27),(153.82±5.04),(901.95±39.30),(109.23±1.18)μg/mg。在细胞试验中,BAPs质量浓度达200μg/mL时,相较于模型组,对所有测定抗氧化指标均产生显著性(P<0.05)影响,细胞存活率显著提高13.80%,细胞内活性氧(ROS)水平下降40.63%,丙二醛(MDA)含量下降11.92 nmol/mg prog,超氧化物歧化酶(SOD)活力和过氧化氢酶(CAT)活力分别提高23.48 U/mg prot 15.71 U/mg prot。BAPs能有效抵抗RAW 264.7细胞产生的氧化应激反应,为开发荞麦多肽为天然抗氧化剂提供理论依据。

To improve the bioavailability of buckwheat protein,buckwheat antioxidant peptides(BAPs) was isolated from the fermentation hydrolysate of buckwheat by using Sephadex G-25 and reversed phase-high performance liquid chromatography(RP-HPLC),and its structure and composition were preliminarily analyzed.The H_2O_2-induced RAW 264.7macrophages cell model of oxidative stress was further established to evaluate the antioxidant activity of BAPs.The re sults showed that the BAPs obtained after separation by RP-HPLC has a purity of 96.25% and a molecular weight of5.8 ku.Vitamin C equivalent antioxidant capacity(VCEAC) was used to indicate that DPPH·,ABTS+·,·OH scavenging ability and reducing power of BAPs were(12.12 ±0.27),(153.82 ±5.04),(901.95 ±39.30),(109.23 ±1.18) μg/mg,respectively.In the RAW 264.7 macrophage oxidative stress model,BAPs with the concentration of 200 μg/mL had a significant(P<0.05) effect on all measured antioxidant indicators compared with the model group,which significantly(P<0.05) promoted the viability of oxidatively damaged cells by 13.80%,reduced the levels of intracellular reactive oxygen species(ROS) by 40.63% and malondialdehyde(MDA) by 11.92 nmol/mg prog.Meanwhile,the activities of superoxide dismutase(SOD) and catalase(CAT) were increased by 23.48 and 15.71 U/mg prot at this concentration,respectively.The above results showed that BAPs can effectively inhibit the oxidative stress response of RAW 264.7 macrophages,which could provide a theoretical basis for the development of buckwheat peptides as natural antioxidants.