银柴胡的生物碱成分及其抗炎活性研究

Study on anti-inflammatory activities of alkaloids from Stellaria dichotoma

目的研究银柴胡Stellaria dichotoma的生物碱成分及其抗炎活性。方法利用硅胶柱色谱、MCI柱色谱和反相制备液相色谱等手段进行分离纯化,并通过1H-NMR、13C-NMR等波谱技术进行结构鉴定。采用脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7炎症反应模型,探究银柴胡中卡巴林生物碱类对巨噬细胞经LPS刺激后产生的炎症因子[肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)]的影响,评价其抗炎活性。结果从银柴胡正丁醇萃取物中分离得到9个卡巴林生物碱,分别鉴定为gelebolineA(1)、gelebolineB(2)、cordysininE(3)、3,10-dihydroxy-β-carboline(4)、6-hydroxy-3-methoxycarbonyl-β-carboline (5)、 1-methyl-3-(2-hydroxypropan-2-yl)-2-(5-methoxy-9H-β-carbolin-1-yl)-cyclopentanol (6)、2-aldehydetetra-hydroharmine(7)、pegaharmine J(8)、pegaharmine K(9)。抗炎活性检测表明,化合物1～9对LPS刺激的巨噬细胞IL-6炎症因子均具有较强的抑制作用。结论所有化合物均为首次从该植物中分离得到,其中化合物1、2对炎症因子IL-6的抑制作用最强,其IC50值最小仅为5.21μg/m L,该研究首次发现银柴胡的抗炎成分。

Objective To study the alkaloids and its anti-inflammatory effect of Stellaria dichotoma. Methods The chemical constituents of S. dichotoma were isolated by different column chromatographic techniques, including silica gel, MCI, and reversed phase prep-HPLC. The structures of these compounds were identified by a comprehensive analysis of the spectroscopic data. LPS-induced RAW264.7 inflammatory cell model was established to evaluate the effect of carboline alkaloids in S. dichotoma on proinflammatory factors(TNF-α and IL-6) of RAW264.7 inflammatory cells. Results Nine compounds were isolated from Stellaria dichotoma. The compounds were identified as geleboline A(1), geleboline B(2), cordysinin E(3), 3,10-dihydroxy-β-carboline(4), 6-hydroxy-3-methoxycarbonyl-β-carboline(5), 1-methyl-3-(2-hydroxypropan-2-yl)-2-(5-methoxy-9 H-β-carbolin-1-yl)-cyclopentanol(6), 2-aldehydetetra-hydroharmine(7), pegaharmine J(8), pegaharmine K(9). The anti-inflammatory activity assay showed that these nine compounds all had strong inhibited effect on production of IL-6 in RAW264.7 inflammatory cell model. Conclusion All the compounds are isolated from this plant for the first time. Activity assay showed that compound 1 and 2 presented better inhibition for proinflammatory factor IL-6, the minimum IC50 is 5.21 μg/mL, and it is the first time to discover the anti-inflammatory constituents of S. dichotoma.