淫羊藿多糖醇沉工艺优化及其对巨噬细胞释放NO的影响
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摘要
旨在优化淫羊藿多糖醇沉条件并研究不同浓度乙醇醇沉的淫羊藿多糖对巨噬细胞释放NO的影响。采用乙醇沉淀法从淫羊藿多糖提取物中获得淫羊藿多糖,在单因素试验基础上,选择浓缩比、乙醇浓度、乙醇体积倍数、醇沉时间为自变量,采用L9(34)的正交试验设计,优化淫羊藿多糖最佳醇沉条件。用浓度为400、500、600、700、800mL/L乙醇醇沉的淫羊藿多糖,采用体外培养法,测定淫羊藿多糖对巨噬细胞释放NO的影响。结果表明,淫羊藿多糖最佳醇沉条件为浓缩比为1∶2,乙醇浓度为800mL/L,乙醇体积倍数为4倍,醇沉时间为12h;每个乙醇浓度醇沉的淫羊藿多糖均具有良好的活性,500mL/L乙醇醇沉的多糖活性最好。结果显示,淫羊藿多糖对巨噬细胞具有良好的刺激作用。
To optimize the alcohol precipitation conditions of Epimediumpolysaccharide and study the effect on nitric oxide release from macrophages,the extraction technology was used by using ethanol precipitation technology.On the basis of single factor test results,concentration ratio,ethanol concentration,volume ratio of ethanol,time of alcohol precipitation were selected as independent variables.Four factors and three levels of the orthogonal experimental design were used to optimize extraction of Epimedium polysaccharide.The effect of Epimedium polysaccharide on the release of NO from macrophages was determined.Epimediumpolysaccharide concentrations were set as 400,500,600,700,800 mL/L alcohol.The result showed that optimal alcohol precipitation conditions of Epimediumpolysaccharide:concentration ratio was1∶2,ethanol concentration was 800mL/L,volume ratio of ethanol was 4,time of alcohol precipitation was12 h.The polysaccharide of Epimedium with each ethanol concentration had good activity.500mL/L alcohol precipitation was the best.The result showed that Epimedium polysaccharide had a good stimulating effect on macrophages.
To optimize the alcohol precipitation conditions of Epimediumpolysaccharide and study the effect on nitric oxide release from macrophages,the extraction technology was used by using ethanol precipitation technology.On the basis of single factor test results,concentration ratio,ethanol concentration,volume ratio of ethanol,time of alcohol precipitation were selected as independent variables.Four factors and three levels of the orthogonal experimental design were used to optimize extraction of Epimedium polysaccharide.The effect of Epimedium polysaccharide on the release of NO from macrophages was determined.Epimediumpolysaccharide concentrations were set as 400,500,600,700,800 mL/L alcohol.The result showed that optimal alcohol precipitation conditions of Epimediumpolysaccharide:concentration ratio was1∶2,ethanol concentration was 800mL/L,volume ratio of ethanol was 4,time of alcohol precipitation was12 h.The polysaccharide of Epimedium with each ethanol concentration had good activity.500mL/L alcohol precipitation was the best.The result showed that Epimedium polysaccharide had a good stimulating effect on macrophages.
引文
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[2]桑野,李晓光,张昌浩,等.国内植物多糖的研究及发展趋势[J].吉林医药学院学报,2017,38(6):455-457.
[3]安晓娟,冯琳,宋红平,等.植物多糖的结构分析及药理活性研究进展[J].中国药学杂志,2012,47(16):1271.
[4]何余堂,潘孝明.植物多糖的结构与活性研究进展[J].食品科学,2010,31(17):493-496.
[5]李翠丽,王伟,张英,等.中药多糖提取、分离纯化方法的研究进展[J].中国药房,2016,27(19):2700-2703.
[6]舒任庚,蒋跃平,蔡永红.植物多糖的提取分离方法探讨[J].中国药房,2011,22(11):1052-1055.
[7]蒋俊,罗珍,刘艳芳,等.醇沉条件对猴头菌多糖得率和品质的影响[J].食用菌学报,2011,18(2):57-61.
[8]顾欢欢,褚秀玲,苏建青.响应面法优化纤维素酶辅助提取淫羊藿多糖工艺[J].中国兽药杂志,2017,51(8):52-58.
[9]尹美珍,李世普,袁琳,等.小鼠腹腔巨噬细胞的分离培养与鉴定[J].武汉大学学报(医学版),2006,27(2):203-205.
[10]李海涛,肖丹,屈学民,等.小鼠腹腔巨噬细胞的分离培养[J].现代生物医学进展,2008,8(4):638-639.
[11] Lin H Y,Juan S H,Shen S C,et al.Inhibition of lipopolysaccharide-induced nitric oxide production by flavonoids in RAW264.7macrophages involves heme oxygenase-1[J].Biochem Pharmacol,2003,66:1821-1832.