几种人参属中药提取物的制备及体外抗卵巢癌活性比较

目的:从人参、西洋参、竹节参、三七这4个人参属中药中提取成分,对其抗卵巢癌活性进行筛选、比较。方法:人参、西洋参、竹节参和三七分别用水和乙醇热回流提取制备相应的水提物和醇提物;上述药材70%乙醇水提取物经大孔树脂柱色谱分别制备得到总皂苷和相应的原人参二醇型皂苷和原人参三醇型皂苷部位。用噻唑蓝(MTT)法对这些提取物和部位在卵巢癌HEY细胞进行活性筛选,并用碘化丙啶(PI)染色检测提取物对细胞周期分布的影响,进一步用免疫印迹法(Western blot)检测了其对细胞周期蛋白表达的影响。划痕实验检测了提取物对细胞迁移能力的影响。结果:各提取物都表现出不同程度地抑制卵巢癌HEY细胞增殖活性,其中西洋原人参二醇型皂苷(PDSPQ)、竹节参总皂苷(TSPJ)、人参醇提物(EEPG)、人参总皂苷(TSPG)、人参原人参二醇型皂苷(PDSPG)对HEY细胞的增殖抑制比较明显,抑制率分别为45.59%,45.78%,50.48%,46.98%,64.36%,PDSPQ,TSPG,PDSPG可以不同程度的下调周期蛋白依赖性激酶(CDK)4,CDK6,细胞周期蛋白D1(cyclin D1)的表达,但对细胞迁移能力没有明显影响。结论:含有原人参二醇型皂苷的组分的抗卵巢癌HEY细胞活性比其他部位更明显,其机制可能与下调细胞周期相关蛋白CDK4,CDK6,cyclin D1的表达有关。

Extraction of Panax Chinese Medicines and Their Anti-cancer Effects on HEY Ovarian Cancer Cells

Objective: To extract effective components from Panax ginseng, P. quinquefolium, P. japonicas and P. notoginseng, and compare their inhibitory activities against ovarian cancer HEY cells. Method: Water and ethanol extracts of P. ginseng, P. quinquefolium, P. japonicas and P. notoginseng were prepared by reflux extraction with water and ethanol. Total saponins and their panaxadiol saponin (PDS) and panaxtrol saponin (PTS) fractions were prepared from 70% ethanol water extracts of above herbs using macroporous resin chromatography. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to detect the activities of these extracts and fractions against HEY ovarian cancer cells. propidium iodide(PI) staining was used to detect the effect of extracts on cell cycle distribution, and western blot was performed to detect their effect on expression of cell cycle related proteins. In addition, scratch assay was conducted to detect the effect of extracts on cell migration. Result: All the indicated extracts showed varied activities against HEY cells. Among them, panaxadiol saponin of P. quinquefolium (PDSPQ), total saponin of P. japonicas (TSPJ), ethanol extract of P. ginseng (EEPG), total saponin of P. ginseng (TSPG), and panaxadiol saponin of of P. ginseng(PDSPG) showed relative higher inhibitory rate against HEY cells, with 45.59%, 45.78%, 50.48%, 46.98% and 64.36%, respectively. Further, PDSPQ, TSPG, and PDSPG could down-regulate the expressions of CDK4, CDK6, and cyclin D1 to different degrees, but had no significant effect on the migration of HEY cells. Conclusion: Fractions with PDS were more effective on inhibiting HEY cells growth, and the mechanism may be associated with down-regulating the protein expressions of CDK4, CDK6, and cyclin D1.