人参属植物高温蒸制前后人参皂苷含量的变化和细胞毒活性研究

目的研究人参属植物人参Panax ginseng、三七P. notoginseng和西洋参P. quinquefolium高温蒸制前后主要人参皂苷的含量变化,并测定高温蒸制前后样品对4株肿瘤癌细胞的细胞毒活性。方法采用HPLC建立了测定22种皂苷含量的分析方法,测定这些皂苷在人参属植物及其高温蒸制品中的含量。用MTT法测定人参属植物及其高温蒸制品对4株人类癌细胞(人类骨髓癌HL-60细胞、肝癌SMMC-7721细胞、肺癌A-549细胞、乳腺癌SK-BR-3细胞)的细胞毒活性。结果从人参、三七和西洋参及高温蒸制后的样品中鉴定出22个皂苷,包括人参皂苷Rg1、Re、Rb1、Rc、Rb2、Rd、Rk3、Rh4、Rk1、Rg5、Rb3、Rh3、Rk2,20(S)-人参皂苷Rh1、20(R)-人参皂苷Rh1、三七皂苷Fc、三七皂苷R1、绞股蓝皂苷IX、20(S/R)-三七皂苷Ft1、20(S/R)-人参皂苷Rg3、人参皂苷Rs3、人参皂苷Rh2。人参皂苷Rg1、Re、Rb1、Rc、Rb2和Rd为人参的主要成分;人参皂苷Rg1、Re、Rb1和Rd为西洋参的主要成分;人参皂苷Rg1、Re、Rb1、Rd和三七皂苷R1为三七的主要成分,高温蒸制后这3种植物的主要成分全部转化为人参皂苷Rk3、Rh4、Rk1、Rg5和20(S/R)-人参皂苷Rg3,在高温蒸制后的三七中还检测到20(S)-人参皂苷Rh1和20(R)-人参皂苷Rh1。三七皂苷Fc、人参皂苷Rb3和绞股蓝皂苷IX为三七茎叶的主要成分,高温蒸制后转化为另外8个主要成分20(S/R)-三七皂苷Ft1,20(S/R)-人参皂苷Rg3、Rs3、Rk1、Rg5,20(S/R)-人参皂苷Rh2、Rh3和Rk2。细胞毒活性结果显示,高温蒸制三七的细胞毒活性比高温蒸制人参和高温蒸制西洋参强,高温蒸制三七的细胞毒活性比三七的活性强,说明高温蒸制后三七的细胞毒活性增强。结论人参、西洋参和三七经过高温蒸制后原来的主要成分基本消失,随之转化为其他主要成分,高温蒸制三七的细胞毒活性最强。

Variation of ginsenosides in raw and processed ginsengs (Panax sp.) and their cytotoxicities

Objective To study the changes in the content of the major ginsenosides of the raw roots and the steaming processed ones of three Panax plants, e.g. Renshen (Panax ginseng), Sanqi (Panax notoginseng) and Xiyangshen (Panax notoginseng) and evaluate the cytotoxicities of raw roots and the steaming processed ones of three Panax plants against four cancer cell lines. Methods An HPLC method was established to qualify the amounts of 22 saponins. The amounts of these saponins in the raw roots and the steaming processed ones were quantified. The cytotoxicities of the raw roots and the steaming processed ones against four cancer cell lines (human myeloid leukemia HL-60, hepatocellular carcinoma SMMC-7721, lung cancer A-549, and breast cancer SK-BR-3 cell lines) were evaluated by MTT methods. Results From them, 22 saponins, including ginsenosides Rg₁, Re, Rb₁, Rc, Rb₂, Rd, Rk₃, Rh₄, Rk₁, Rg₅, Rb₃, Rh₃, Rk₂, 20(S)-and 20(R)-ginsenoside Rh₁, notoginsenosides Fc and R₁, gypenoside IX, 20(S/R)-notoginsenoside Ft₁, 20(S/R)-ginsenosides Rg₃, Rs₃, and Rh₂ were identified. Among them, ginsenosides Rg₁, Re, Rb₁, Rc, Rb₂and Rd, ginsenoside Rg₁, Re, Rb₁and Rd, and ginsenoside Rg₁, Re, Rb₁, Rd and notoginsenosides R₁were found to be the major saponins in the raw roots of P. ginseng, P. quinquefolium, and P. notoginseng, respectively. However, after steaming process, they all changed to be ginsenosides Rk₃, Rh₄, Rk₁, Rg₅ and 20(S/R)-ginsenosides Rg₃. In case of the steaming processed roots of P. notoginseng, two more ginsenosides, 20(S)-ginsenoside Rh₁and 20(R)-ginsenoside Rh₁ were also detected at the same time. Three saponins, notoginsenoside Fc, ginsenoside Rb₃ and gypenoside IX were detected to be the major saponins in the leaves and stems of P. notoginseng, and after steaming process, most of them were changed to other eight saponins, 20(S/R)-notoginsenosides Ft₁, Rg₃, and Rs₃, ginsenosides Rk₁, Rg₅, and 20(S/R)-ginsenoside Rh₂, Rh₃, Rk₂. It was noted that the processed P. notoginseng was more cytotoxic than processed P. ginseng and processed P. quinquefolium. The activity of cytotoxicities of the processed P. notoginseng was better than that of raw P. notoginseng. It means that after steaming process, P. notoginseng become more cytotoxic against four cancer cell lines. Conclusion The main ingredients of three Panax plants disappeared and transformed to others. The cytotoxicities of processed P. notoginseng were stronger than others.