红凉伞根化学成分及抗炎活性研究

Chemical constituents and their anti-inflammatory activities from the roots of Ardisia crenata var. bicolor

关键词:  红凉伞根, 化学成分, 分离纯化, 结构鉴定, 岩白菜素, 抗炎活性

Key words:  Ardisia crenata var. bicolor root, chemical constituents, isolation and purification, structure identification, bergenin, anti-inflammatory activity

摘要:

红凉伞(Ardisia crenata var. bicolor)为贵州苗族常用药,具有清喉利咽、消肿止痛、祛风除湿等功效。为研究红凉伞根化学成分及体外抗炎活性,该文采用硅胶柱色谱、Sephadex LH-20凝胶柱色谱、ODS反相柱色谱及半制备HPLC等方法对红凉伞根70%乙醇提取物进行系统性研究,结合NMR、MS等现代波谱技术分析及文献对比进行化合物结构鉴定。采用脂多糖(LPS)诱导的小鼠RAW 264.7巨噬细胞的NO生成模型,评价化合物的抗炎活性。结果表明:(1)从红凉伞根70%乙醇提取物中分离得到10个化合物,分别鉴定为11-O-没食子酰岩白菜素(1)、11-O-(4-O-甲基没食子酰)岩白菜素(2)、11-O-香草酰岩白菜素(3)、6-O-(4-羟基苯甲酰基)岩白菜素(4)、11-丁香酰岩白菜素(5)、11-O-(3',4'-二甲基没食子酸)-岩白菜素(6)、去甲氧基岩白菜素(7)、micractinin A(8)、monomethyl olivetol(9)、邻苯二甲酸二丁酯(10),其中化合物4、8、9为首次从紫金牛属中分离得到。(2)体外抗炎活性实验结果表明,化合物1-4对小鼠RAW 264.7细胞的NO生成具有明显的抑制作用(P<0.01),在20 μmol·mL^-1浓度下,NO抑制率分别为67.09%、66.50%、59.83%、36.47%。该研究结果丰富了红凉伞根的化学成分,明确了其部分抗炎活性物质基础,为其药材资源的开发利用提供了理论依据。

Abstract:

Ardisia crenata var. bicolor is a common medicine used by Miao Minority in Guizhou, which has the effects of clearing throat and benefiting pharynx, reducing swelling and relieving pain, dispelling wind and dehumidifying. In order to study the chemical constituents and anti-inflammatory activities of A. crenata var. bicolor roots, the 70% ethanol extact was separated and purified by silica gel chromatography, Sephadex LH-20 gel column chromatography, ODS reverse column chromatography and semi-preparative HPLC. The structures of the compounds were identified by spectral data of NMR, MS and published literatures. Using lipopolysaccharide(LPS)-activated RAW 264.7 cell line model in vitro, compounds 1-4 were evaluated for the inhibition against nitric oxide(NO)production. The results were as follows:(1)Ten compounds were isolated from the 70% ethanol extract and identified as 11-O-galloylbergenin(1), 11-O-(4-O-methylgalloyl)bergenin(2), 11-O-vanilloylbergenin(3), 6-O-(4-hydroxy benzoyl)bergenin(4), 11-syringyl bergenin(5), 11-O-(3',4'-dimethylgalloyl)-bergenin(6), demethoxybergenin(7), micractinin A(8), monomethyl olivetol(9), and dibutyl phthalate(10). Among them, compounds 4, 8, 9 were obtained from Ardisia for the first time.(2)The results of anti-inflammatory activities in vitro showed that compounds 1-4 could significantly inhibit NO release in RAW 264.7 cells(P<0.01), and the inhibition rates of compounds at the concentration of 20 μmol·mL^-1 were 67.09%, 66.50%, 59.83%, 36.47%, respectively. This study enriches the chemical constituents of A. crenata var. bicolor roots, clarifies the material basis of its anti-inflammatory activities, verifies the scientificity of its traditional usage, and provides new insight and scientific evidence for its rational development and utilization of the medicinal resources.