北豆根化学成分及其抗炎活性研究

Chemical constituents from the rhizome of Menispermum dauricum and their anti-inflammatory activities

关键词:  防己科, 北豆根, 化学成分, 结构鉴定, 抗炎活性

Key words:  Menispermaceae, rhizome of Menispermum dauricum, chemical constituents, structure identification, anti-inflammatory activity

摘要:

北豆根,防己科蝙蝠葛( Menispermum dauricum )的干燥根茎,是重要的中药材。为阐明北豆根的药效物质基础,该研究利用硅胶柱层析、大孔吸附树脂及高压制备HPLC等色谱技术对北豆根化学成分进行系统分离,采用NMR等波谱技术对化合物进行结构鉴定,并通过测定化合物对脂多糖(LPS)诱导的巨噬细胞RAW264.7释放 NO和IL-6炎症因子的含量评价其抗炎活性。结果表明:(1)从北豆根甲醇提取物中分离鉴定了15个化合物,分别鉴定为对羟基苯甲醛(1)、香草酸(2)、丁香醛(3)、2-羟基-1-(4-羟基-3,5-二甲氧基苯基)-1-丙酮(4)、对羟基苯乙酸甲酯(5)、2-(4-羟苯基)-硝基乙烷(6)、对羟基苯乙腈(7)、邻苯二甲酸二丁酯(8)、fragransin B₂(9)、7-hydroxy-3,6-dimethoxy-1,4-phenanthraquinone(10)、棕榈酸(11)、花生酸(12)、 β-谷甾醇(13)、 β-豆甾醇(14)、胡萝卜苷(15)。其中,化合物 4 -7、9、12为首次从防己科植物中分离得到,化合物1、3-11、14为首次从蝙蝠葛属植物中分离得到。上述化合物包括苯酚类、木脂素类、菲醌类、脂肪酸类及甾醇类。(2)化合物12在25、50 μg·mL^-1浓度下可以显著抑制LPS诱导的RAW264.7细胞NO和IL-6的释放,具有潜在的抗炎作用。该研究结果极大地丰富了北豆根的化学多样性,明确了其抗炎活性的活性成分,为药用植物资源的合理利用提供了有效参考。

Abstract:

Bei-dou-gen, the dried rhizome of Menispermum dauricum from the Menispermaceae family, is an important Chinese medicinal material. In order to clarify the pharmacodynamic substance basis, the methanol extract of the rhizome in M. dauricum was systematically isolated and purified using various chromatographic methods and the structures of isolated compounds were identified. The potential anti-inflammatory activity of obtained compounds from M. dauricum was measured in vitro. In this study, the chemical constituents were separated via silica gel column chromatography, macroporous adsorption resin and preparative HPLC, and their structures were determined on the basis of MS, ¹H-NMR, ¹³C-NMR and other spectroscopic data analysis, as well as comparison with relevant literatures. Meanwhile, the anti-inflammatory activities of against NO and IL-6 production from LPS-induced RAW264.7 cells of the chemical components were evaluated in vitro. The results were as follows:(1)Fifteen compounds were isolated from the rhizome of M. dauricum, and identified as p-hydroxybenzaldehyde(1), vanillic acid(2), syringaldehyde(3), 2-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone(4), methyl 4-hydroxyphenylacetate(5), 2-(4-hydroxyphenyl)-1-nitroethane(6), 4-hydroxyphenylacetonitrile(7), dibutyl phthalate(8), fragransin B₂ (9), 7-hydroxy-3,6-dimethoxy-1,4-phenanthraquinone(10), palmitic acid(11), arachidic acid(12), β-sitosterol(13), β-stigmasterol(14)and daucosterol(15). Among them, compounds 4-7, 9 and 12 were isolated from Menispermaceae for the first time, while compounds 1, 3-11 and 14 were first reported from Menispermum genus. The above compounds were all non-alkaloids components, including phenols, lignans, phenanthraquinones, fatty acids and sterols.(2)Anti-inflammatory assay in vitro showed that Compound 12 could significantly inhibited releases of NO and IL-6 induced by LPS from RAW264.7 cells at the concentrations of 25 and 50 μg·mL^-1, indicating potential anti-inflammatory activity. The results of this study enriches the chemical diversity of the rhizome of M. dauricum and clarifies the active ingredients of its anti-inflammatory activity. It provides an effective reference for the rational utilization of medicinal plant resources.