目的 优选分离丹参二萜醌最佳的高速逆流色谱(HSCCC)溶剂体系,明确丹参二萜醌的体外抗肿瘤活性。方法 采用CO₂超临界萃取法制备丹参总二萜醌,采用UPLC测定丹参酮II_A和隐丹参酮在不同溶剂系统上下相中的峰面积,计算分配系数(K)值及K值的比值(α),确定最佳溶剂体系。CKK-8法观察丹参二萜醌对人肝癌(QGY-7703)、肺癌(PC9、A549)、胃癌(MKN-45、HGC-27)、结肠癌(HCT116)、骨髓瘤(U266、RPMI8226)、乳腺癌(MCF-7)等9种人源肿瘤细胞株的抑制作用。结果 HSCCC溶剂系统为石油醚-醋酸乙酯-甲醇-水(12:8:13:7)时,丹参酮II_A和隐丹参酮能得到较好的分离,丹参二萜醌的得率为8.65%。丹参二萜醌体外对人肝癌、肺癌、胃癌、结肠癌、骨髓瘤、乳腺癌等9种人源肿瘤细胞均有一定的抑制作用,其中对人肺癌PC9细胞及人乳腺癌MCF-7细胞的抑制作用最为显著。结论 确定的溶剂体系对丹参二萜醌有效部位的分离效果可靠,建立的HSCCC制备方法操作简单,可作为高效快速分离纯化丹参二萜醌的分离制备方法。丹参二萜醌在体外具有良好的抗肿瘤活性。

Objective To optimize the solvent system of high-speed counter-current chromatography (HSCCC) for diterpenoidtanshinone separation and define it's antitumor activity in vitro. Methods Total diterpenoidtanshinone was made by CO₂ supercritical fluid extraction (SFE), UPLC was applied to determining the peak area of tanshinone II_A and cryptotanshinone in different solvents up or under phase, and the partition coefficient K values of them were calculated. CKK-8 was used to observe the inhibitory effects of diterpenoidtanshinone on human liver cancer (QGY-7703), lung cancer (PC9, A549), gastric cancer (MKN-45, HGC-27), colon cancer (HCT116), myeloma (U266, RPMI8226), and breast cancer (MCF-7). Results The best solvent system for HSCCC was petroleum ether-ethyl acetate-methanol-aqua (12:8:13:7). The yield of diterpenoidtanshinone was 8.65%. Diterpenoidtanshinone has good effect of antitumor in vitro especially on human PC9 cell. Conclusion The selected solvent system is suitable for diterpenoidtanshinone separation by HSCCC, and the established HSCCC method is reliable and easy for operating. Diterpenoidtanshinone has good antitumor effect in vitro.

高等学校博士学科点专项科研基金资助课题(博导类)(20123322110001);浙江省重大科技专项计划项目(2012C13017-1);中国博士后科学基金(2014M561791)