白英抗肿瘤活性成分研究

【作者】 臧雅丽；

【导师】 王建农；

【作者基本信息】 中国中医科学院 ， 中药学， 2008， 硕士

【摘要】 白英,别名“白毛藤”,来源于茄科植物白英Solanum lyratum Thunb.的全草,味苦、辛,性微寒,归肝、胆经,具有清热解毒、祛风化痰、利湿退黄、抗癌的功能,为常用抗癌中草药。研究内容分为白英的组分分离,白英组分对小鼠移植性肿瘤作用的体内抗肿瘤实验,白英有效部位的化学研究,有效部位及单体的体外抗肿瘤活性筛选,白英单味药质量标准的制定五个部分。通过大孔树脂柱对白英醇提液进行粗分离,富集到白英的两类主要化学成分-白英总甾体皂苷（SLTG）部位和白英总甾体生物碱（SLTA）部位。对SLTG部位、SLTA部位及二者的配伍进行小鼠移植性肿瘤S₁₈₀肉瘤及艾氏腹水瘤（EAC）体内抗癌活性研究。结果显示SLTA部位对小鼠移植性肿瘤S₁₈₀肉瘤及艾氏腹水瘤（EAC）均有很好的抑瘤效果,尤以中剂量组的抑瘤效果最为明显;SLTG部位只有中剂量组对小鼠移植性肿瘤S₁₈₀肉瘤有一定的抑制作用;两部位配伍的抑瘤效果反而弱于将二者分开时的抑瘤效果,因此SLTA部位为白英抗肿瘤的主要药效部位,对该部位进行系统的化学研究。运用硅胶柱色谱法、氧化铝柱色谱法及薄层色谱法等方法,对白英抗肿瘤主要药效部位的化学成分进行系统研究,从该部位分离到6个单体化合物。经理化性质、薄层鉴别、NMR图谱、MS谱及文献比对确定6个化合物分别为雅姆皂苷元、薯蓣皂苷元、澳洲茄碱、澳洲茄二烯、木糖基苦茄碱及木糖基澳洲茄边碱,其中澳洲茄碱的得率最高。利用MTT法对SLTA部位及得率最高的澳洲茄碱单体进行体外抗肿瘤活性筛选,选取的细胞瘤株为人肝癌细胞株BEL-7402、人肺癌细胞株A-549、人直肠癌细胞株HCT-8。结果SLTA部位在剂量为50μg·mL^-1时,对A-549细胞株的生长有一定的抑制作用;澳洲茄碱对上述三种细胞瘤株没有抑制作用,对BEL-7402细胞株生长反而有微弱的促进作用。利用比色法以澳洲茄碱为对照品测定SLTA部位中总生物碱的含量并进行方法学考察,结果所建立的方法可靠、可行,该部位以澳洲茄碱计总生物碱含量为51.18%;薄层色谱法以雅姆皂苷元、澳洲茄碱为对照品对SLTA部位进行定性鉴别,从而为白英单味药材制定了部分质量标准。更多还原

【Abstract】 The whole plant of Solarium lyratum Thunb is called "baimaoteng" in clinical application which property is cool, flavor is bitter and goes back to liver and guts meridian. It has effect on heat-clearing and detoxicating, dispelling wind-evil and dissipate phlegm, jaundice with damp-heat pathogen, anticancer etc. It’s often used as a remedy for various cancers.The content of this text composed of five parts which are separation of chemial composition of Solarium lyratum Thunb, the antitumous effect of Solarium lyratum Thunb’s chemial composotion on portability tumous of mouse in vivo, studying on the chemical constituent of main antitumous chemial composotion, competent screening in main antitumous chemial composition and compound by experiment in vitro, formulating part quality specification for Solarium lyratum Thunb.Through macroporous resin to enrich two main constituent of Solarium lyratum Thunb’s alcohol extraction to get Solanum Lyratum Total Glycosides （SLTG） composition and Solanum Lyratum Total Alkaloid （SLTA） composition. Then by pharmacodynamic action research in vivo to find main antitumous chemial composotion, this text chose portability tumous of mouse which are S₁₈₀ and EAC malignant cell strains as indexes to decide the antitumous effect of SLTG, SLTA, SLTG and SLTA compatibility, total extraction of Solanum Lyratum. The result show that SLTA composition has obvious antitumous effect on S₁₈₀ and EAC, especially the mid dose experiment group has obvious antitumous effect; the mid dose group of SLTG composition has antitumous effect, other experimental groups have no antitumous effect. It’s worth noting that the rate of antitumous of SLTG composition and SLTA composition compatibility experimental groups are lower than SLTG composition and SLTA composition separating experimental groups. From the pharmacodynamic action research in vivo SLTA is the main antitumous composition, so do chemical research about it. Through silica gel column chromatograph, aluminium oxidehe column chromatograph and thin layer chromatography （TLC） obtain 6 compounds from SLTA, by the aid of physico-chemical property, TLC, NMR spectrum, MS spectrum and literature to definite their constitutional formula. They are yamogenin, diosgenin, solasonine, solasodiene, xylosyl-solamarine, xylosyl-solamarine, among them solasonine has a highly yield. Utilizing MTT method to do antitumous research in vitro about SLTA and compounds, this text chose BEL-7402, A-549 and HCT-8 malignant cell strains of person. The result show that when SLTA’s concentration is 50ug·mL^-1, it has effect on A-549, but other three low concentration have no effect; Solasonine has no effect on three malignant cell strains; on the contrary, it hastenes the growth of BEL-7402 malignant cell strains. Then formulate part quality standard for Solarium lyratum Thunb. Use colorimetric method to determine the Steroid alkaloid content of SLTA with solasonine as standard substance, the result is 51.18%. Use TLC to formulate qualitation discrimination with solasonine and yamogenin as standard substance.更多还原