瞿麦正丁醇部位抗肿瘤活性成分筛选  

Screening of Antineoplastic Constituents in n-butanol Fraction of Dianthus superbus

作　　者：张建超[1] 余建清[2] 张方蕾[1] 张磊[1] 江银[1] 刘焱文[1] 

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]武汉大学药学院,湖北武汉430072

出　　处：《中国现代中药》2015年第4期326-330,共5页Modern Chinese Medicine

基　　金：国家自然科学基金资助项目(30970294)

摘　　要：目的:研究瞿麦(Dianthi herba)正丁醇部位中具有抗肿瘤活性的化合物。方法:用乙醇提取瞿麦全草中的化学成分,正丁醇萃取后,在用四甲基偶氮唑盐(MTT)法检测Hep G2细胞活性的筛选下,用硅胶凝胶柱色谱以及半制备HPLC分离纯化,得到对肿瘤细胞株的细胞毒活性成分VI3-15-1-1,并通过LC-MS、1H、13C NMR,结合二维核磁图谱,推测出该活性成分为积雪草酸(Asiatic acid,2α,3β,23α-三羟基-熊果-12-烯-28-酸)。结果:通过MTT法分别测定瞿麦正丁醇部位硅胶柱色谱分离得到的8个组分,其中第6个组分(30-33组分)对Hep G2细胞增殖抑制率最高,为28.69%;进一步分离第6个组分得到的VI3-15组分对Hep G2细胞增殖抑制率为91.14%。从VI3-15分离得到的化学成分为积雪草酸,其对Hep G2、SSMC-7721、SK-HEP-1及Bel-7402细胞株的半数抑制率(IC50)分别为55.4、48.9、55.3、58.8μg·m L-1,在体外具有较好地抑制肿瘤细胞增殖的活性。结论:确定瞿麦正丁醇部位抗肿瘤活性成分为五环三萜类化合物积雪草酸。Objective： To study the antineoplastic constituent in Dianthus superbus. Methods： The dried aerial parts of D. superbus were extracted with EtOH（95% ）, the extracts were combined and concentrated under reduced pressure to afford the extraction which was suspended in water and extracted with petroleum ether, EtoAc, n-BuOH. The n-BuOH solution was concentrated to give the n-BuOH soluble fraction which was submitted to silica gel, sephadex LH-20, preparative HPLC and afforded active compound. The cytotoxieity of each fraction against HepG2 cells was investigated by MTF method. On the basis of LC-MS、^1H,^13C NMR and combined with two-dimensional NMR spectra, the chemical structure of active compound has been elucidated as asiatic acid（2α, 3β, 23α-trihydroxyurs-12-en-28-oic acid）. Result： The in vitro anticancer activities of eight fractions separated from n-BuOH fraction by sihca gel column chromatography were investigated by MTT assay. Among them, the most efficient fraction inhibited for HepG2 cells was VI fraction（ 30-33 ）with inhibition ratio 28. 69%. The VI3-15 fraction separated from VI showed the inhibition ratio 91.14% on HepG2 cells. Asiatic acid purified from VI3-15 fraction showed inhibitory effects on HepG2, SSMC-7721, SK-HEP-1 and Bel-7402 cells, with half-inhibitory concentrations（IC50）at 55.4, 48.9, 55.3, 58.8 μg·mL^-1, respectively. Asiatic acid showed good antineoplastic activities. Conclusion： The antineoplastic compound of n-butanol fractionfrom D. superbus is asiatic acid.

关 键 词：瞿麦 抗肿瘤活性 积雪草酸 

分 类 号：R979.1[医药卫生—药品]