怀菊花总黄酮富集纯化工艺优化及其抗炎活性、成分组成研究  

Optimization of enrichment and purification process for total flavonoids from Huai Chrysanthemum and their anti-inflammatory activity,component composition

作　　者：邓晓颜 王小兰[1,2] 李孟 张莉[1,2] 李玉贤 郑晓珂[1,2] 冯卫生 DENG Xiao-yan;WANG Xiao-lan;LI Meng;ZHANG Li;LI Yu-xian;ZHENG Xiao-ke;FENG Wei-sheng(College of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Provincial Engineering and Technology Center for Chinese Medicine Development,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州450046

出　　处：《中成药》2023年第9期2815-2821,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(8210142016);河南省科技攻关项目(212102310345);河南省重大科技专项项目(171100310500);河南省高等学校重点科研项目(21B360003);河南省中医药科学研究专项项目(20-21ZY2148)。

摘　　要：目的优化怀菊花总黄酮富集纯化工艺,并考察其抗炎活性、成分组成。方法在单因素试验基础上,以乙醇解吸体积分数、乙醇解吸体积、水除杂体积为影响因素,解吸率为评价指标,响应面法优化富集纯化工艺。采用CCK-8法检测总黄酮对ox-LDL诱导HUVECs细胞的增殖作用,ELISA法检测IL-1β、IL-6、TNF-α水平。HPLC法分析总黄酮成分组成。结果最佳条件为乙醇解吸体积分数80%,乙醇解吸体积2 BV,水除杂体积4 BV,解吸率为84.93%,纯度为52.71%。与ox-LDL组比较,不同剂量(25、50、100μg/mL)总黄酮组HUVECs细胞增殖率升高(P<0.05,P<0.01),IL-1β、IL-6、TNF-α水平降低(P<0.05,P<0.01)。总黄酮含有木犀草苷、芹菜素-7-O-葡萄糖苷、香叶木素-7-O-葡萄糖苷、木犀草素、金合欢素-7-O-葡萄糖苷、芹菜素、香叶木素、金合欢素、异绿原酸A。结论该方法简便高效,可用于富集纯化具有抗炎活性的怀菊花总黄酮。AIM To optimize the enrichment and purification process for total flavonoids from Huai Chrysanthemum,and to investigate their anti-inflammatory activity,component composition.METHODS With ethanol desorption concentration,ethanol desorption volume and water impurity removal volume an influencing factors,desorption rate as an evaluation index,the enrichment and purification process was optimized by response surface method based on single factor test.CCK-8 method was applied to detecting the proliferation effects of total flavonoids on(ox-LDL)-induced HUVECs cells,and ELISA method was adopted in the detection of IL-1β,IL-6,TNF-αlevels.The component composition of total flavonoids was analyzed by HPLC.RESULTS The optimal conditions were determined to be 80%for ethanol desorption concentration,2 BV for ethanol desorption volume,and 4 BV for water impurity removal volume,the desorption rate and purification rate were 84.93%and 52.71%,respectively.Compared with the ox-LDL group,the different doses(25,50,100μg/mL)of total flavonoids groups demonstrated increased purification rates of HUVECs cells(P<0.05,P<0.01),and decreased IL-1β,IL-6,TNF-αlevels(P<0.05,P<0.01).Total flavonoids contained sluteolin,apigenin-7-O-glucoside,geranyllinin-7-O-glucoside,luteolin,acacia-7-O-glucoside,apigenin,geranyllinin,acacia and isochlorogenic acid A.CONCLUSION This simple and efficient method can be used for the enrichment and purification of total flavonoids from Huai chrysanthemum with anti-inflammatory activity.

关 键 词：怀菊花总黄酮 富集纯化工艺 抗炎活性 成分组成 响应面法 CCK-8 ELISA HPLC 

分 类 号：R284.2[医药卫生—中药学]