摘要
本研究建立脂多糖诱导的巨噬细胞炎症模型,结合层析法对土牛膝抗炎活性成分进行分离,并用NMR、MS以及与对照品比较,对得到的化合物进行结构鉴定。结果表明粗毛牛膝醇提取物抗炎效果最佳,其反相层析50%、70%及100%甲醇洗脱物抗炎活性高和细胞毒性低,从50%-70%甲醇洗脱物分离鉴定出9种单体化合物:β-蜕皮甾酮(1)、牛膝甾酮(2)、水龙骨甾酮B(3)、N-反式阿魏酰酪胺(4)、N-顺式阿魏酰酪胺(5)、N-顺式阿魏酰-3-甲氧基酪胺(6)、N-反式阿魏酰-3-甲氧基酪胺(7)、竹节参皂苷Ⅳa(8)、5,2′-二甲氧基-6-甲氧甲基-7-羟基-异黄酮(10);100%甲醇洗脱物鉴定出主要成分竹节参皂苷Ⅰ(9)。在LPS+25μM单体浓度下,化合物抗炎活性由强到弱为4>1>5>2>6>9>10>3>7>8。反相液相色谱测定结果表明,野生牛膝和柳叶牛膝(两年生)地下根茎β-蜕皮甾酮含量在温度高季节含量高,8月份达到最高,分别为0.914±0.016和1.412±0.038 mg/g;两年生粗毛牛膝随季节变化规律不明显。具有抗炎活性的阿魏酰酪胺类生物碱首次从土牛膝中分离鉴定。
The inflammation macrophages(RAW 264.7) model induced by lipopolysaccharide(LPS) with nitric oxide(NO) release as an index was established to investigate the anti-inflammatory components in different varieties of Tuniuxi and column chromatography was used to isolate the active compounds from Achyranthes aspera Linnacus methanol extract.The chemical structures of the active components were deduced based on the nuclear magnetic resonance(NMR) and mass spectrometry(MS) data analysis and comparing with reference substances.The methanol extracts of A.aspera showed the best anti-inflammatory effect,furthermore,its 50%,70% and 100% methanol elutes featured low cytotoxicity and high anti-inflammatory activity.Nine pure compounds isolated from the 50% and 70% methanol elutes were identified as β-ecdysterone(1),inokosterone(2),polypodine B(3),N-trans-feruloyl-tyramine(4),N-cis-feruloyl-tyramine(5),N-cis-feruloyl-3-methoxytyramine(6),N-trans-feruloyl-3-methoxytyramine(7),chikusetsusaponin IVa(8),5,2′-dimethoxy-6-(methoxymethyl)-7-hydroxy-isoflavonol(10),meanwhile,oleanolic acid 28-O-β-D-glucopyranosyl ester(9) was elucidated as the main component in its 100% methanol elute.At the concentration of LPS + 25 μM,the anti-inflammatory activity order was 4 > 1 > 5 > 2 > 6 > 9 > 10 > 3 > 7 > 8.The β-ecdysterone content in the underground rhizomes of wild A.bidentata Blume and A.longifolia(Makino) Makino changed obviously with seasons and reached the highest level in August at 0.914±0.016 and 1.412±0.038 mg/g,respectively.However,there is no obvious seasonal variation observed in the A.aspera.Moreover,feruloyl tyramine alkaloids with anti-inflammatory activity were firstly reported from A.aspera in this research.
作者
欧阳文
罗懿钒
李震
王雄龙
张云坤
周华荣
唐纯玉
李顺祥
OUY ANG Wen;LUO Yi-fang;LI Zhen;WANG Xiong-long;ZHANG Yun-kun;ZHOU Hua-rong;TANG Chun-yu;LI Shun-xiang(School of Phrmacy,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Era Sunshine Pharmaceutical Co.,Ltd.,Yongzhou 410116,China;Hunan Food and Drug Vocational College;Hunan Proince Engineering Research Center of Bioaetive Substance Discovery of TCM,Changsha 410208,China;Hunan Engineering Technology Research Center for anti.infection Chinese Medicine;Hunan Era Sunshine Collaborative R&D Center for Post adoctoral Sudies,Yongzhou 410116,China)
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2020年第7期1171-1181,共11页
Natural Product Research and Development
基金
湖南省教育厅优秀青年基金(18B251)
湖南省食品药品监督管理局食品药品安全科技项目(湘食药科R201806)
永州市科技创新及应用研究项目(永财企指[2019]2号)
湖南中医药大学中药学一流建设学科(校行科字[2018]3号)。
关键词
土牛膝
抗炎活性
分离鉴定
β-蜕皮甾酮
含量测定
Achyranthes aspera
anti-inflammatory activity
isolation and identification
β-ecdysterone
content determination
