摘要
目的:以金银花为研究对象,构建不同萃取阶段中绿原酸的纯化工艺并评估其抗炎活性。方法:采用石油醚、乙酸乙酯、正丁醇等有机溶剂对金银花70%乙醇提取物进行萃取,并采用高效液相色谱法检测不同萃取阶段绿原酸的含量。用脂多糖(LPS)诱导小鼠巨噬细胞(RAW264.7)建立炎症模型,用MTT法测细胞存活率,一氧化氮(NO)试剂盒检测NO浓度,并评估不同含量绿原酸的抗炎活性。结果:乙酸乙酯层、正丁醇层、第一次上清、第二次上清、70%醇提浸膏及脱脂浸膏绿原酸含量分别为26.17%、11.86%、9.27%、7.80%、6.95%和6.52%。不同萃取层获得的绿原酸均能显著降低细胞NO分泌量(p<0.05),NO生成抑制率依次为悬浮物层(95.56%)>石油醚层(94.95%)>第一次沉淀(69.85%)>乙酸乙酯层(41.73%)>第二次沉淀(35.95%)>醇提浸膏(20.66%)。结论:随着指标成分绿原酸含量的提高,其抗炎活性逐渐变小,金银花发挥最大抗炎活性组分为石油醚层。
Objective:To construct a purification process for chlorogenic acid in different extraction stages and evaluate its anti-inflammatory activity in Lonicera japonica Thunb.as the research object.Methods:70%ethanol extract of Lonicera japonica Thunb.is extracted by petroleum ether,ethyl acetate,n-butanol and other organic solvents,and the content of chlorogenic acid in different extraction stages is detected by high performance liquid chromatography.Lipopolysaccharide(LPS)induced mouse macrophages(RAW264.7)to establish an inflammation model,cell viability is measured by MTT method,nitric oxide(NO)kit is used to detect NO concentration,and the anti-inflammatory activity of different contents of chlorogenic acid is evaluated.Results:The chlorogenic acid contents of ethyl acetate layer,n-butanol layer,first supernatant,second supernatant,70%alcohol extraction extract and defatted extract are 26.17%,11.86%,9.27%,7.80%,6.95%and 6.52%,respectively.The chlorogenic acid obtained from different extraction layers can significantly reduce the NO secretion of cells(p<0.05),and the inhibition rate of NO formation is in the suspended solids layer(95.56%)>petroleum ether layer(94.95%)>the first precipitation(69.85%)>the ethyl acetate layer(41.73%)>the second precipitation(35.95%)>alcohol extraction extract(20.66%).Conclusion:With the increase of the content of chlorogenic acid,the anti-inflammatory activity of the index ingredient chlorogenic acid gradually decreased,and the maximum anti-inflammatory activity component of Lonicera japonica Thunb.is the petroleum ether layer.
作者
柯春山
吴磊
谢传奇
徐志勇
熊伟
卞海生
KE Chunshan;WU Lei;XIE Chuanqi;XU Zhiyong;XIONG Wei;BIAN Haisheng(Institute of Applied Chemistry,Jiangxi Academy of Sciences,Nanchang 330096,China;Jinguli Flower Planting Farmers’Professional Cooperative in Aktao County,Aktao 845551,China)
出处
《生物化工》
CAS
2024年第5期43-47,60,共6页
Biological Chemical Engineering
基金
江西省科技对口支援项目(20240BDB29006)。
关键词
金银花
绿原酸
萃取溶剂
抗炎活性
Lonicera japonica Thunb.
chlorogenic acid
extraction
anti-inflammatory activity
