北五味子多糖提取工艺优化及其对LPS刺激巨噬细胞线粒体膜电位的保护作用

Optimization of Extraction Technology of Schisandra chinensis Polysaccharide and Its Protective Effect on LPS Induced Mitochondrial Membrane Potential in Macrophages

目的:提取北五味子多糖,研究其体外抗氧化活性及对脂多糖(LPS)诱导的巨噬细胞线粒体膜电位的保护作用。方法:通过热水浸提法及四因素三水平正交分析法L9(3^4)提取北五味子多糖,对比北五味子多糖与维生素E聚乙二醇琥珀酸酯(VE-TPGS)的体外抗氧化活性,利用LPS构建巨噬细胞损伤模型,通过活性氧(ROS)、JC-1细胞染色实验研究北五味子多糖对ROS的抑制作用及对细胞线粒体膜电位的保护作用。结果:北五味子多糖提取工艺为:料液比1∶40 g/mL,提取次数3次,提取时间3.0 h,提取温度为90℃,在此条件下多糖最高得率为10.72%,北五味子多糖总糖含量为84.28%±2.03%,葡萄糖含量为30.26%±1.12%;北五味子多糖清除DPPH自由基的IC50值为602.52μg/mL,清除羟自由基的IC50值为34.24μg/mL,北五味子多糖1600μg/mL时,总抗氧化能力达到(0.28±0.01)mmol/mg。细胞实验表明北五味子多糖能够降低LPS诱导巨噬细胞的活性氧的水平,抑制细胞线粒体膜电位下降。结论:北五味子多糖在体外具有较强的抗氧化活性,能够抑制LPS诱导巨噬细胞中活性氧水平进而发挥其对线粒体膜电位的保护作用。

Objective:Studying the antioxidant activity of Schisandra chinensis polysaccharides in vitro and its protective effect on mitochondrial membrane potential of macrophages induced by lipopolysaccharide(LPS).Methods:Schisandra chinensis polysaccharides was extracted by hot water extraction method and four-factor three-level orthogonal analysis method L9(34).Compared the antioxidant activity of Schisandra chinensis polysaccharides with vitamin E polyethylene glycol succinate(VE-TPGS)in vitro.The model of macrophage induced injury by LPS to study the inhibitory effect of Schisandra chinensis polypeptide on ROS and the protective effect on cell mitochondrial membrane potential by reactive oxygen species(ROS)and JC-1 cell staining.Results:The extraction process of polysaccharide from Schisandra chinensis was material-liquid ratio 1∶40 g/mL,the extraction times 3 times,the extraction time 3.0 h,the extraction temperature 90℃.Under this condition,the highest yield was 10.72%.The total sugar contents of Schisandra chinensis polysaccharide was 84.28%±2.03%,the glucose contents was 30.26%±1.12%,and the IC50 value of DPPH free radical scavenged was 602.52μg/mL,hydroxyl radical scavenged was 34.24μg/mL by Schisandra chinensis polysaccharide.When the Schisandra chinensis polysaccharide was 1600μg/mL,the total antioxidant capacity reaches 0.28±0.01 mmol/mg.Cell experiments showed that Schisandra chinensis polysaccharide could reduce the level of reactive oxygen species and inhibit the decreasing of mitochondrial membrane potential after LPS induced macrophages.Conclusion:The Schisandra chinensis polysaccharide had strong antioxidant activity in vitro,and would inhibit LPS-induced reactive oxygen levels and exert its protective effect on mitochondrial membrane potential in macrophages.