民族药刺梨根茎化学成分及其抗炎活性研究

Chemical constituents and their anti-inflammatory activities from rhizome of ethnic medicine Rosa roxburghii

关键词:  民族药, 刺梨根茎, 化学成分, 分离鉴定, 抗炎活性

Key words:  ethnic medicine, rhizome of Rosa roxburghii, chemical constituents, isolation and identification, anti-inflammatory activity

摘要:

民族药刺梨根茎在贵州少数民族地区有着广泛的应用,为验证其化学成分的抗炎活性,该文以民族药新鲜刺梨根茎为原料,采用硅胶柱色谱、Sephadex LH-20柱色谱等方法对其根茎的化学成分进行分离纯化,通过理化性质和NMR等波谱数据鉴定化合物的结构; 采用脂多糖(LPS)诱导的小鼠巨噬细胞RAW 264.7作为炎症模型,考察刺梨根茎化学成分对巨噬细胞经LPS刺激后产生的NO炎症因子的影响,评价其抗炎活性。结果表明:(1)从刺梨根茎乙醇提取物中共分离获得15个化合物,结构分别鉴定为刺梨苷(1)、野蔷薇苷(2)、蔷薇酸(3)、β-D-glucopyranosyl-(2a→1b)-2a-O-β-L-arabinopyranosyl-(2b→1c)-2b-O-β-L-arabinopyranosyl-(2c→1d)-2c-O-β-L-arabinopyranosyl-(2d→1e)-2d-O-β-L-arabinopyranosyl-(2e→1f)-2e-O-β-L-arabinopyranoside(4)、儿茶素(5)、3-O-methylellagic acid-4'-O-β-D-xylopyranoside(6)、3-O-methylellagic acid-4'-O-α-L-rhamnopyranoside(7)、委陵菜酸(8)、桦木酸(9)、spinosic acid(10)、arjunic acid(11)、 β-谷甾醇(12)、β-胡萝卜苷(13)、α-tocopherol(14)、正二十六烷(15)。其中化合物4、6、7首次从该植物中分离得到。(2)对其中的化合物1-7进行体外抗炎活性实验,结果发现化合物1-7对LPS诱导的小鼠巨噬细胞RAW 264.7释放的NO均有明显抑制作用,且呈剂量依赖关系; 化合物1-7在抗炎作用上表现出较好活性,其IC₅₀值分别为25.07、24.56、17.65、9.87、16.67、40.83、34.98 μmol·L^-1(阳性对照地塞米松为22.46 μmol·L^-1 ),其中化合物3、4、5的活性优于地塞米松。该研究结果阐明了刺梨根茎中的三萜类、鞣花酸类、黄酮类和寡糖类化合物是其抗炎作用的主要有效成分,并验证了刺梨根茎的民间抗炎功效。

Abstract:

To study the anti-inflammatory of chemical constituents from the rhizome of ethnic medicine Rosa roxburghii. The chemical constituents of fresh R. roxburghii rhizome were studied by separation and purification technology, and their structures were identified through physicochemical properties and spectral data. The inflammatory model of RAW 264.7 induced by lipopolysaccharide(LPS)in vitro was established. The cell viability was detected by MTT assay, the release of inflammatory factor NO in the cell culture medium was detected by Griess method and the anti-inflammatory activity of each compound was evaluated through the kit. The results were as follows:(1)Fifteen compounds were isolated and identified as kaji-ichigoside F1(1), rosamultin(2), euscaphicacid(3), β-D-glucopyranosyl-(2a→1b)-2a-O-β-L-arabinopyranosyl-(2b→1c)-2b-O-β-L-arabinopyranosyl-(2c→1d)-2c-O-β-L-arabinopyranosyl-(2d→1e)-2d-O-β-L-arabinopyranosyl-(2e→1f)-2e-O-β-L-arabinopyranoside(4), catechin(5), 3-O-methylellagic acid-4'-O-β-D-xylopyranoside(6), 3-O-methylellagic acid-4'-O-α-L-rhamnopyranoside(7), tormentic acid(8), betulinic acid(9), spinosic acid(10), arjunic acid(11), β-sitosterol(12), β-daucosterol(13), α-tocopherol(14)and n-hexacosane(15). Compounds 4, 6 and 7 were isolated from R. roxburghii for the first time.(2)The results of in vitro anti-inflammatory activity showed that compounds 1-7 significantly inhibited LPS induced NO production from RAW 264.7 in a dose-dependent manner, anti-inflammatory activities of compounds 1-7 were showed better, and the IC₅₀ values were 25.07, 24.56, 17.65, 9.87, 16.67, 40.83 and 34.98 μmol·L^-1, respectively(dexamethasone as positive control 22.46 μmol·L^-1 ), among which compounds 3, 4 and 5 were slightly better than dexamethasone. The results indicate that triterpenoids, ellagic acids, flavonoids and oligosaccharides from R. roxburghii are the main effective components and verify its anti-inflammatory effects in folk application.