克瘤丸组成药物提取物对人肺癌细胞的抑制作用

The Inhibition of Medicine Composition Extracts from Keliu Pills on Human Lung Cancer Cells

目的研究克瘤丸组成药物提取物对人肺癌细胞A549的抑制作用,优化精简克瘤丸的处方组成。方法采用MTT法考察黄芪总皂苷、斑蝥超临界萃取物、虫类药乙醇提取物、山豆根总生物碱和黄芪、山豆根水提物等克瘤丸5个提取物对A549细胞增殖的抑制作用;选用五因素二水平正交实验设计,分析克瘤丸5个提取物对A549细胞增殖的不同效应,判断其是否存在交互作用。结果细胞毒实验结果表明克瘤丸中的斑蝥萃取物、虫类药醇提物、黄芪总皂苷和山豆根总生物碱等4种提取物对A549的IC50值分别是0.330、1.041、1.961、2.015mg/mL。其中斑蝥萃取物和虫类药醇提物的抑制作用48h达到峰值,存在量效和时效关系。正交实验结果显示,虫类药醇提物在抑制A549细胞增殖中起主要作用,斑蝥萃取物和黄芪总皂苷次之。方差分析表明黄芪总皂苷和虫类药醇提物、斑蝥萃取物和虫类药醇提物存在交互作用。结论虫类药醇提物、斑蝥萃取物和黄芪总皂苷是克瘤丸发挥作用的必要成分,可精简为3者的配伍方。

OBJECTIVE To study the inhibitory effects of five medicine composition extracts from Keliu Pills on human lung adenocarcinoma A549 cells and streamline the prescription.METHODS MTT assay was used to investigate antiproliferative effects of five extracts（total astragalosides,supercritical fluid extracts of cantharis,alcohol extract of insect,total alkaloids from tonkinensis and aqueous extract of astragalus and tonkinensis）on A549 cells.Five factors two levels orthogonal experiment design was used to determine the different antiproliferative effects of five extracts against A549 cells and to explore the interaction between each extract.RESULTS Cytotoxic experiment results showed that IC50 values of supercritical fluid extracts of cantharis,alcohol extract of insect drugs,total astragalosides and total alkaloids from tonkinensis were 0.330,1.041,1.961,1.041mg/mL,respectively.The inhibitory rates of supercritical fluid extracts of cantharis,alcohol extract of insect drugs reached the peak at 48 h,with a concentration-effect and time-effect relationship.Orthogonal experiment showed that alcohol extract of insect drugs played a main role in antiproliferation against A549 cells.Supercritical fluid extracts of cantharis and total astragalosides followed.Analysis of variance showed that interaction effects were found between the total astragalosides and alcohol extract of insect drugs,as well as supercritical fluid extracts of cantharis and alcohol extract of insect drugs.CONCLUSION The alcohol extract of insect drugs,supercritical fluid extracts of cantharis and total astragalosides are necessary for Keliu Pills,which can be streamlined for the compatibility of these three extracts.