三脉紫菀化学成分的分离及其抗补体和抗炎活性的研究

Isolation of chemical ingredients from Aster ageratoides and their anticomplement and anti-inflammatory activities

目的活性导向分离鉴定三脉紫菀Aster ageratoides中具有抗补体活性的主要成分,探讨其中活性较好的化合物抗补体主要作用靶点以及抗炎活性。方法采用减压硅胶柱、凝胶sephadexLH-20柱色谱法、中压制备和半制备液相法,对三脉紫菀醋酸乙酯部位进行具有抗补体活性的化学成分分离,其结构通过1H-NMR及13C-NMR进行鉴定;采用红细胞溶血法对分离得到的化学成分进行抗补体活性及其作用靶点筛选;采用脂多糖(LPS)刺激小鼠单核巨噬细胞RAW264.7细胞对化合物进行抗炎活性的研究。结果分离得到14个化合物,分别鉴定为齐墩果酸(1)、槲皮素(2)、山柰酚(3)、3,5,7,3′-四羟基-4′-甲氧基黄酮(4)、山柰酚-7-O-α-L-吡喃鼠李糖苷(5)、槲皮素-3-O-β-D-吡喃葡萄糖苷(6)、山柰酚-3-O-α-L-鼠李糖苷(7)、槲皮素-3-O-α-L-鼠李糖苷(8)、山柰素-3-O-β-D-葡萄糖苷(9)、山柰酚-3-O-β-D-葡萄糖苷(10)、山柰酚-7-O-β-D-葡萄糖苷(11)、山柰酚-3-O-β-D-葡萄糖-7-O-β-D-葡萄糖苷(12)、山柰素-3-O-α-L-鼠李糖-(1→6)-β-D-葡萄糖苷(13)、芦丁(14)。抗补体实验结果表明,14个化合物都具有一定的抗补体活性,并呈现较好的构效关系。抗补体靶点研究结果表明,化合物1作用于补体系统的C1q、C5、C9组分,化合物2作用于补体系统的C1q、C2、C5、C9。抗炎活性研究表明化合物能明显抑制一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的释放,且能明显抑制诱导型一氧化氮合酶(i NOS)和环氧化酶-2(COX-2)的表达。结论分离得到14个化合物,其中化合物1、4、6、9、12、13为该植物中首次发现。这14个化合物具有一定程度的抗补体活性,且化合物2具有明显的抗炎活性。

Objective To isolate and identify chemical ingredients with anti-complement activity from Aster ageratoides and investigate the key targets and anti-inflammatory activities of obtained compounds with good anti-complement activity. Methods Using silica gel column, sephadex LH-20 column, Medium Pressure Liquid Chromatography system, and Semi-preparative HPLC,chemical ingredients that displayed anti-complement activity were isolated. Their chemical structures were identified by 1 H-NMR and13 C-NMR and their anti-complement activities and targets were investigated by erythrocyte hemolysis in vitro. In addition, using LPS-stimulated RAW264.7 cells, we investigated the anti-inflammatory activity of compound 2. Results A total of 14 compounds were obtained from A. ageratoides and identified as oleanolic acid(1), quercetin(2), kaempferol(3), 3,5,7,3′-tetrahydroxy-4′-methoxyflavone(4), kaempferol-7-O-α-L-rhamnopyranoside(5), quercetin-3-O-β-D-glucopyranoside(6), kaempferol-3-O-α-Lrhamnoside(7), quercetin-3-O-α-L-rhamnoside(8), kaempferide-3-O-β-D-glucopyranoside(9), kaempferol-3-O-β-Dglucopyranoside(10), kaempferol-7-O-β-D-glucopyranoside(11), kaempferol-3-O-β-D-glucopyranoside-7-O-β-D-glucopyranoside(12), kaempferide-3-O-α-L-rhamnoside-(1→6)-β-D-glucopyranoside(13), and rutin(14). They all exhibited anti-complement activity to some certain degree and good structure-activity relationship. The targets of compounds 1 and 2 were C1 q, C5, and C9 and C1 q, C2, C5, and C9, respectively. The anti-inflammatory experiments indicated that compound 2 exhibited a significant biological activity, which significantly suppressed the release of NO, TNF-α, and IL-6 and expressions of iNOS and COX-2. Conclusion A total of 14 compounds were obtained and they all displayed anticomplement activity, of which compounds 1, 4, 6, 9, 12, and 13 are firstly discovered in A. ageratoides. Compound 2 exhibited a significant anti-inflammatory activity.