摘要
目的研究半枝莲和白花蛇舌草药对中总黄酮富集的工艺条件及体外抗氧化和抑菌作用。方法以总黄酮的含量、收率为考察指标,筛选D101型大孔吸附树脂富集药对中总黄酮的吸附和洗脱参数,采用DPPH法及FRAP法来评价药对中总黄酮的抗氧化活性。采用“双层平板打孔法”评价药对中总黄酮对金黄色葡萄球菌和大肠杆菌的抑制作用,二倍稀释法测定总黄酮的最低抑菌浓度(MIC),通过绘制生长曲线、测定菌体胞内蛋白的泄漏初步探讨其抑菌机制。结果D101型大孔吸附树脂富集的工艺条件为:7 BV上样液(总黄酮浓度为2.52 mg·mL^(-1))以1.0 mL·min^(-1)速度进行吸附,用5.8 BV纯化水洗脱,10 BV 81%乙醇以3 mL·min^(-1)速度洗脱,药对中总黄酮的平均含量可达26.40%;与富集前比较,含量提高了将近3倍。抗氧化作用表明:富集黄酮的IC50为26.7μg·mL^(-1),粗黄酮的IC50为69.7μg·mL^(-1),对DPPH均有良好的清除作用,富集黄酮和粗黄酮的FRAP值较高,都具有良好的总抗氧化能力。抑菌作用表明:药对中总黄酮对金黄色葡萄球菌有抑制作用,对大肠杆菌无明显的抑制作用。粗黄酮、富集黄酮对金黄色葡萄球菌的MIC均为3.75 mg·mL^(-1)。药对中总黄酮可能是通过破坏细胞壁或膜的结构,导致细胞膜通透性增加,从而影响细菌正常的生长和繁殖。结论富集总黄酮的工艺条件简单、可行,药对中总黄酮具有一定的抗氧化及抑菌作用,且均呈浓度依赖关系。
OBJECTIVE To study enrichment parameters and antioxidant and antibacterial effects in vitro of total flavonoids from Scutellariae barbatae and Hedyotis diffusa.METHODS Taking the content and yield of total flavonoids as indexes,the absorption and elution parameters of D101 macroporous resin was screened.The antioxidant activity of total flavonoids was evaluated by DPPH and FRAP methods.The effect of total flavonoids on Escherichia coli and Staphylococcus aureus was studied by using Double-layer plate punching method.MIC values were determined by double dilution method.Antibacterial mechanism was studied by drawing growth curves and determining protein leakage.RESULTS The conditions of concentration technology were as follows:the sample volume of 7 BV(equivalent to concentration of total flavonoids of 2.52 mg·mL^(-1))at a velocity of 1.0 mL·min^(-1) was absorbed,and eluted with purified water of 5.8 BV,and then eluted with 81%ethanol of 10 BV at a speed of 3 mL·min^(-1).The average content of total flavonoids in the herb pair can attain 26.40%and is nearly 3 times higher than before.Enriched and crude flavonoids had good antioxidant activity(IC50 is 26.7μg·mL^(-1) and IC50 is 69.7μg·mL^(-1),respectively).Enriched and crude flavonoids had certain antibacterial effect(MIC=3.75 mg·mL^(-1)).The total flavonoids from the herb pair could influence the growth and reproduction of bacteria by destroying the composition of wall or membrane of cell.CONCLUSION The process conditions for the enrichment of total flavonoids are simple and feasible,and the results show that the total flavonoids have certain antioxidant and antibacterial activities in a concentration-dependent manner.
作者
王桂玲
费洪荣
赵雪梅
赵莹
WANG Guiling;FEI Hongrong;ZHAO Xuemei;ZHAO Ying(Shandong First Medical University&Shandong Academy of Medical Sciences,Taian,Shandong,271016 P.R.China)
出处
《华西药学杂志》
CAS
CSCD
2021年第3期268-275,共8页
West China Journal of Pharmaceutical Sciences
基金
山东省高等学校科技计划项目(J16LM03)
泰安市科技发展计划(2018NS0146)
山东省医药卫生科技发展计划(2018WS134)。
关键词
半枝莲
白花蛇舌草
药对
总黄酮
大孔吸附树脂
富集
抗氧化
抑菌
Scutellaria barbata D.Don
Hedyotis diffusa Willd
Herb pair
Total flavonoids
Macroporous adsorption resin
Enrichment
Antioxidant effect
Antibacterial effect
作者简介
王桂玲,女,吉林长春,硕士,从事天然药物活性成分的研究工作。Email:wgl20050711@sina.com。
