正交设计优选丹参-人参组分抗肝癌配伍研究

目的:优选丹参-人参活性组分抗肝癌优化配伍并对其作用机制作初步研究。方法:以人肝癌细胞SMMC-7721为研究对象,人正常肝细胞L-O2为对照,采用CCK-8法以对肝癌细胞增殖抑制和正常肝细胞保护作用为筛选指标,正交设计优选丹参-人参活性组分抗肝癌有效组合,确定优化配伍;应用实时细胞分析技术(RTCA DP)检测丹参-人参组分优化配伍对肝癌细胞SMMC-7721和正常肝细胞L-O2增殖的影响,并进行时效关系考察;采用Annexin V/PI双染法经高内涵细胞成像系统(HCS)检测丹参-人参组分优化配伍对肝癌细胞SMMC-7721凋亡的影响。结果:丹参-人参组分配伍抗肝癌SMMC-7721细胞的优化组合为丹参总酚酸、人参总皂苷和人参多糖,其配伍剂量分别为10,10,5 mg·L-1;优选的丹参-人参组分配伍能显著抑制肝癌细胞的增殖,呈现时间依赖性,而对正常肝细胞的增殖抑制作用不明显;丹参-人参组分优化配伍能诱导肝癌细胞凋亡,对正常肝细胞的凋亡无显著影响。结论:丹参-人参组分优化配伍具有潜在的特异选择性抗肝癌作用,其机制与抑制肝癌细胞增殖和诱导细胞凋亡相关。

Optimization of Effective Component Formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on Hepatoma Carcinoma Using Orthogonal Design

Objective: To optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on hepatoma carcinoma, and to reveal its antitumor mechanisms. Method: Human hepatoma cell SMMC-7721 was used as object and human normal liver cell line L-O2 was used as control in this study. The inhibiting effect on SMMC-7721 and the protective effect on L-O2 were performed by CCK-8 assay, and the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma was optimized by using the orthogonal design method. The effect on SMMC-7721 and L-O2 viabilities were assayed by using Real-time cell assay and the time-effect relationship was analyzed. Cell apoptosis was measured by high content screening using Annexin V/PI double staining. Result: The results demonstrated that the contents of total salvianolic acid, total saponins Ginseng Radix et Rhizoma and ginseng polysaccharide doses were 10, 10, 5 mg·L^-1, respectively, in the optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma . Furthermore, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could significantly decrease the SMMC-7721 cell viability and induce SMMC-7721 cell apoptosis, while there was no significant difference on the normal liver cell. Conclusion: The component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma has potentially specific and selective anti-hepatoma carcinoma effect. Its mechanism may be related to the inhibition of hepatoma cell proliferation and induction of cell apoptosis.