为了研究硒化牡丹籽粕多糖的抗氧化活性,水提醇沉法提取牡丹籽粕粗多糖,离子交换层析制备牡丹籽粕多糖(peony seed dreg polysaccharides,PSDP)PSDPⅠ、PSDPⅡ及PSDP Ⅲ。硝酸-亚硒酸钠法对PSDP Ⅲ进行硒化修饰得Se-PSDP Ⅲ。采用体外抗氧化实验,评价Se-PSDP Ⅲ对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基和羟自由基(·OH)的清除作用及对DNA氧化损伤的抑制作用。并以巨噬细胞RAW264.7建立H2O2诱导的氧化损伤细胞模型,探讨Se-PSDP Ⅲ在细胞水平的抗氧化能力。结果表明:Se-PSDP Ⅲ具有较好的体外清除自由基的能力和抑制DNA氧化损伤的能力。细胞模型法评价结果显示,Se-PSDP Ⅲ可以降低H2O2诱导的巨噬细胞RAW264.7内活性氧(reactive oxygen species,ROS)的生成。此外,Se-PSDP Ⅲ可以升高细胞内超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的活力。因此,硒化牡丹籽粕多糖作为潜在的抗氧化物,可用于功能产品的开发。
To investigate the antioxidant activity of selenium-containing peony seed dreg polysaccharides,the crude polysaccharides were separated by water extraction and alcohol precipitation,which was further purified by ion-exchange chromatography to obtain main polysaccharides (PSDP I,PSDP II and PSDP III). The nitric acid-sodium selenite (HNO3-Na2SeO3) method was used to obtain Se-PSDP III. In vitro antioxidant experiment was used to evaluate the scavenging effect of Se-PSDP Ⅲ on 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals and hydroxyl free radicals (·OH) and inhibition of DNA oxidative damage. In addition,macrophage RAW264.7 was used to establish a H2O2-induced oxidative damage cell model to explore the antioxidant capacity of Se-PSDP Ⅲ at the cellular level. The results demonstrated that Se-PSDP III showed a good ability to scavenge free radicals in vitro and inhibit DNA oxidative damage. The cell model evaluation results showed that Se-PSDP III could protect RAW264.7 macrophages against H2O2-induced oxidative damage by decreasing reactive oxygen species (ROS) production. Moreover,Se-PSDP III could increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Therefore,Se-PSDP III could be used as potential antioxidants in the development of functional products.
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