薏苡附子败酱散方抗肝癌细胞SMMC-7721有效组分配伍研究

Effective components compatibility of Yiyi Fuzi Baijiang Powder against Hepatoma cells SMMC-7721

目的通过均匀设计方法筛选出薏苡附子败酱散方有效组分抗肝癌细胞最佳配伍比例,并对其各有效组分及最佳配伍进行体外初步作用机制研究。方法采用MTT法测定各有效组分对肝癌细胞SMMC-7721的IC_(50)值,通过U_8(8~5)均匀设计对有效组分配伍进行优化,确定最佳配伍比例;同时测定最佳配伍作用于人正常肝细胞L-02和肝癌细胞SMMC-7721的IC_(50)值;运用流式细胞仪将最佳配伍组及有效组分单用组分别作用于肝癌细胞SMMC-7721,测定细胞周期与凋亡,并用Flowjo及Modifit软件进行分析。结果薏苡附子败酱散方有效组分最佳配伍比例为:败酱草总皂苷-薏苡仁挥发油-败酱草总黄酮-附子总生物碱=0.40∶2.06∶1.06∶1.70。最佳配伍对肝癌细胞SMMC-7721的IC_(50)值比对人正常肝细胞L-02显著降低;与空白对照组比较,各组分单用组及最佳配伍组作用于肝癌细胞SMMC-7721后,细胞凋亡率均增加;细胞分裂周期G_0/G_1期、S期比例升高,G_2/M期比例降低。最佳配伍组作用优于各有效组分单用组。结论通过本实验筛选出的薏苡附子败酱散方有效成分最佳配伍对肝癌细胞SMMC-7721有明显的增殖抑制作用,为该方的进一步研发奠定了基础。

Objective To determine the effective component compatibility of Yiyi Fuzi Baijiang Powder against Hepatoma cells, and to study the external preliminary reaction mechanism for the optimal component compatibility. Methods Effective components were tested to determine the IC50 of Hepatoma cells SMMC-7721 with MTT method. The effective component compatibility against Hepatoma cells SMMC-7721 and its reaction against the human normal hepatocyte L-02 and IC50 were determined by the uniform design U8 （8^5）. Optimal compatibility and effective components against hepatocellular SMMC-7721, the cell cycle and apoptosis were measured by flow cytometer, and analyzed by Flowjo and Modifit software. Results The optimal compatibility ratio was determined with： saponins of patrinavillosajus-coix seed essential oil-total flavonoids of patrinavillosajuss-total alkaloid of mookshood ： 0.40 ： 2.06 ： 1.06 ： 1.70. The best compatibility of liver cancer IC50 was significantly lower than that of the normal liver cells. Compared with the blank group, the cell apoptosis rate was increased after using the single component and the optimal component compatibility recipe for the hepatocellular SMMC-7721. The cell proliferation period G0/G1 and the ratio of S time were increased too; the ratio of GJM time was decreased. The reaction of optimal component compatibility was better than that of the single effective component against Hepatoma cells SMMC-7721. Conclusion The selected effective component compatibility ofYiyi Fuzi Baijiang Powder can obviously inhibit Heoatoma cells SMMC-7721.