云木香二氯甲烷提取物抗肝纤维化活性成分的鉴定

Identification of active anti-hepatic fibrosis ingredients of dichloromethane extract from Saussurea costus

目的 云木香二氯甲烷提取物中抗肝纤维化(HF)活性成分的鉴定。方法 云木香药材的粗粉用95%乙醇回流提取,所得浸膏用水溶解,再用石油醚、二氯甲烷、乙酸乙酯及正丁醇依次萃取得到粗提物;应用硅胶、中压色谱分离凝胶(MCI)、Sephadex LH-20凝胶等柱色谱和制备液相色谱等技术对云木香二氯甲烷萃取部位进行分离纯化,采用核磁共振波谱和质谱进行结构鉴定;采用噻唑蓝(MTT)法测定化合物对人肝星状细胞(HSC)株LX-2的增殖作用影响,评价各化合物的抗HF作用。结果 从云木香二氯甲烷部位中分离到16个化合物,鉴定为木香烯内酯(化合物1)、脱氢木香内酯(化合物2)、β-环木香烯内酯(化合物3)、7α-木香醇(化合物4)、瑞诺木烯内酯(化合物5)、10α-羟基青蒿酸(化合物6)、8α-羟基-11β-11,13-二氢去氢海桑内酯(化合物7)、vlasouliolione B(化合物8)、珊塔玛内酯(化合物9)、草蒿素(化合物10)、4-O-和厚朴酚(化合物11)、2,2-氧双(1,4)-二叔丁基苯(化合物12)、桦木酸(化合物13)、4-羟基-3,5-二甲氧基苯甲醛(化合物14)、5-羟甲基糠醛(化合物15)及茴芹内酯(化合物16);活性测试表明,化合物2、8、9及11可以有效抑制LX-2细胞的增殖,半数抑制浓度(IC50)分别为14.9、51.1、47.2及40.8μmol/L。结论 化合物4、11、12及16为首次从云木香二氯甲烷部位中分离得到,化合物2、8、9及11可能是云木香的主要抗HF活性成分。

Objective To investigate active anti-hepatic fibrosis(HF)ingredients of dichloromethane extract from Saussurea costus.Methods Crude power of Saussurea costus was extracted by refluxing with 95%ethanol,and resulting extract was dissolved in water,then extracted with petroleum ether,dichloromethane,ethyl acetate and n-butanol,respectively,to obtain crude extract.Dichloromethane was isolated and purified by column chromatography technique(including silica gel CC,Middle Chromatogram Isolated Gel(MCI)and Sephadex LH-20)and preparation of liquid chromatography,etc.Its structures were determined by nuclear magnetic resonance and mass spectrometric approach.MTT assay was used to assess the effect of the compounds on cell proliferation of the human hepatic stellate cell(HSC)line LX-2 to evaluate its anti-HF activity.Results Sixteen compounds were isolated from the dichloromethane fraction of Saussurea costus and identified as costunolide(1),dehydrocostuslactone(2),β-cyclocostunolide(3),7α-hydroxycostol(4),reynosin(5),10α-hydroxyl-artemisinic acid(6),8α-hydroxy-11β-11,13-dihydrodehydrocostuslactone(7),vlasouliolione B(8),santamarin(9),artesin(10),4-O-methylhonokiol(11),2,2'-oxybis(1,4)-di-tert-butylbenzene(12),betulinic acid(13),4-hydroxy-3,5-dimethoxy-benzaldehyde(14),5-hydroxymethylfurfural(15),and pimpinellin(16),respectively.Activity assay showed that Compounds 2,8,9,and 11 exhibited the inhibitory effects on LX-2 cell proliferation with IC 50 values of 14.9,51.1,47.2,and 40.8μmol/L,respectively.Conclusion Compounds 4,11,12,and 16 are isolated from Saussurea costus for the first time.Compounds 2,8,9,and 11 might be the main active anti-HF ingredients of dichloromethane extract of Saussurea costus.