杏叶防风的化学成分及抗炎活性研究

Chemical constituents of Pimpinella candolleana and their anti-inflammatory activities

关键词:  杏叶防风,化学成分,分离鉴定,RAW264.7细胞,抗炎活性

Key words:  Pimpinella candolleana, chemical constituents, isolation and identification, RAW264.7 cells, anti-inflammatory activity

摘要:

杏叶防风(Pimpinella candolleana)为贵州苗族习用草药,用于黄疸型肝炎、急性胆囊炎等病症的治疗。为探究杏叶防风的化学成分及其抗炎活性,该研究采用硅胶、凝胶、ODS等色谱技术对杏叶防风全草70%乙醇提取物进行分离纯化,通过NMR、MS等波谱数据鉴定化合物结构,采用脂多糖(LPS)诱导的RAW264.7巨噬细胞作为炎症模型,评价单体化合物的抗炎活性。结果表明:(1)从杏叶防风中分离并鉴定了20个化合物,分别为香草醛(1)、芝麻素(2)、2-甲基-2-羟基-5-甲氧基苯并 [d] 氢化呋喃-3-酮(3)、原儿茶醛(4)、1,5-dihydroxy-2,3-dimethoxyxanthone(5)、异鼠李素(6)、山奈酚(7)、8-羟基-2-甲基色原酮(8)、木犀草素(9)、槲皮素(10)、1-O-β-D-葡萄糖-(2S,3S,4R,8E)-2- [(2'R)-2'-羟基棕榈酰胺]-8-十八烯-1,3,4-三醇(11)、异鼠李素-3-O-β-D-半乳糖苷(12)、异槲皮苷(13)、去甲当药醇苷(14)、木犀草素-6-C-α-L-阿拉伯糖苷(15)、山奈酚-3-O-β-D-半乳糖苷(16)、山奈酚-7-O-β-D-葡萄糖苷(17)、木犀草素-7-O-β-D-葡萄糖苷(18)、异牡荆苷(19)、芦丁(20)。其中,化合物1、3、4、6、7、10、13、16、18、20均为首次从该植物中分离得到。(2)抗炎结果显示,化合物2-10、12、18、19均可显著抑制 LPS 诱导 RAW264.7 细胞NO释放量(P<0.05,P<0.01),其中化合物4、7、10、18在浓度为25 μmol·L^-1时,抑制率分别为57.37%、83.60%、68.16%、81.14%。该研究丰富了杏叶防风的化学成分,明确了黄酮类化合物是其发挥抗炎功效的活性成分,为杏叶防风的进一步研究与开发利用提供了一定的依据。

Abstract:

Pimpinella candolleana is known as Miao ethnic herbal medicine in Guizhou for the treatment of icteric hepatitis, acute cholecystitis and other diseases. To investigate the chemical constituents of P. candolleana and their anti-inflammatory activities, the chemical constituents from the 70% ethanol extract of P. candolleana were separated by silica gel, Sephadex LH-20, Toyopearl HW-40F, Toyopearl HW-40C, ODS and other column chromatography technologies, and their structures were elucidated by extensive spectroscopic analysis such as nuclear magnetic resonance(NMR)and mass spectrum(MS). The inflammatory cell model, built by LPS-induced RAW264.7 macrophage cells, was used to evaluate the anti-inflammatory activity. The results were as follows:(1)Twenty compounds from P. candolleana were isolated and identified of including vanillin(1), sesamin(2), 2-methyl-2-hydroxy-5-methoxy berz(d)hydrofuran-3-one(3), procatechin(4), 1,5-dihydroxy-2,3-dimethoxyxanthone(5), isorhamnetin(6), kaempferol(7), 8-hydroxy-2-methylchromone(8), luteolin(9), quercetin(10), 1-O-β-D-glucopyranosyl-(2S,3S,4R,8E)-2-[(2'R)-2'-hydroxypalmitoylamino]-8-octadecene-1,3,4-triol(11), isorhamnetin-3-O-β-D-galactopyranoside(12), isoquercitrin(13), norswertianolin(14), luteolin-6-C-α-L-arabinoside(15), kaempferol-3-O-β-D-galactopyranoside(16), kaempferol-7-O-β-D-glucopyranoside(17), luteolin-7-O-β-D-glucopyranoside(18), isovitexin(19), rutin(20). Compounds 1, 3, 4, 6, 7, 10, 13, 16, 18, and 20 were obtained from this plant for the first time.(2)The anti-inflammatory results showed that compounds 2-10, 12, 18 and 19 could significantly inhibit the LPS-induced NO content in RAW264.7 cells(P<0.05, P<0.01), and the inhibition rates of compounds 4, 7, 10, and 18 at a concentration of 25 μmol·L^-1 were 57.37%, 83.60%, 68.16%, 81.14%, respectively. Overall, this study enriches the chemical constituents of P. candolleana, and clarifies that flavonoids are the active ingredients in the course of anti-inflammatory, which provides a theoretical reference for further research and exploitation of P. candolleana.