基于BV-2小胶质细胞模型导向分离糙叶五加根的化学成分

Chemical constituents of roots of Acanthopanax henryi isolated based on BV-2 microglia model

目的基于BV-2小胶质细胞模型导向分离糙叶五加Acanthopanax henryi根的化学成分。方法采用脂多糖(lipopolysaccharide,LPS)诱导的BV-2小胶质细胞建立神经炎症模型,以MTT法测定细胞毒性,对糙叶五加根的正己烷、醋酸乙酯、正丁醇提取部位进行活性筛选,确定活性部位,根据筛选结果对最佳活性部位进一步采用柱色谱、D101大孔树脂、半制备液相等方法进行分离纯化,根据理化性质和所得到的光谱数据,鉴定化合物的结构。并对其中得到的部分化合物进行活性验证。结果糙叶五加醋酸乙酯提取物部位的活性最佳,从中分离得到17个化合物,分别鉴定为(+)-芝麻素(1)、松柏醛(2)、acuminatolide(3)、芝麻素酮(4)、10-二十九烷醇(5)、(±)-N-甲基-4-羟基癸酰胺(6)、罗汉松脂素(7)、(1R,2S,5R,6S)-6-(3,4-dihydroxyphenyl)-2-(3,4-methylenedioxyphenyl)-3,7-dioxabicyclo[3,3,0]octane(8)、1-辛烯(9)、胡椒醇(10)、(-)-松脂醇(11)、榕醛(12)、丁香醛(13)、丁香脂醇(14)、(-)-表丁香脂素(15)、原儿茶醛(16)、原儿茶酸(17)。其中分离得到的(+)-芝麻素和丁香醛在细胞耐受浓度(5~80μmol/L)下能减少LPS诱导的BV-2小胶质细胞中一氧化氮(NO)的释放,且呈剂量相关性。结论化合物3~10、12、15首次从五加科植物中分离得到,化合物2~12、15~16首次从糙叶五加中分离得到。糙叶五加根中的化合物芝麻素和丁香醛具有潜在的抗神经炎活性。

Objective To study the chemical constituents and anti-neuritis activity of the root of Acanthopanax henryi.Methods A neuroinflammation model was established using lipopolysaccharide(LPS)-induced BV-2 microglial cells,and cytotoxicity was measured using MTT method.The n-hexane,ethyl acetate,and n-butanol extracts from the roots of A.henryi were screened for activity to determine the active site.Based on the screening results,the optimal active site was further separated and purified using column chromatography,D101 macroporous resin,and semi-preparative solution methods.Based on the physicochemical properties and obtained spectral data,the structure of compound was identified.Results The best activity was observed in the ethyl acetate extract site,from which 17 compounds were isolated and identified as(+)-sesamin(1),coniferaldehyde(2),acuminatolide(3),sesaminone(4),10-nonacosanol(5),rac-N-methyl-4-hydroxydecanamide(6),matairesinol(7),(1R,2S,5R,6S)-6-(3,4-dihydroxyphenyl)-2-(3,4-methylenedioxyphenyl)-3,7-dioxabicyclo[3,3,0]octane(8),1-octene(9),piperitol(10),(−)-pinoresinol(11),ficusal(12),syringaldehyde(13),syringaresinol(14),(−)-episyringaresinol(15),protocatechuyl aldehyde(16),protocatechuic acid(17),respectively.The isolated sesamin and syringaldehyde can reduce the release of NO in BV-2 microglia induced by LPS in a dosedependent manner at cell tolerance concentrations(5—80μmol/L).Conclusion Compounds 3—10,12,15 were isolated from family Araliaceae for the first time.Compounds 3—10,12,15 were isolated from the Acanthopanax Miq.for the first time.Compounds 2—12,15—16 were isolated from A.henryi for the first time.The monomeric compounds sesamin and syringaldehyde in the roots of A.henryi have potential anti-neuroinflammatory activities.