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目的:研究中药两头尖中竹节香附素A、竹节香附皂苷R2及竹节香附皂苷R0的体外抗肿瘤生物活性,比较侧链糖的个数对抗肿瘤活性的影响,并通过检测细胞内活性氧的含量研究其在细胞凋亡过程中的作用。方法:采用体外MTT法,研究竹节香附素A、竹节香附皂苷R2及竹节香附皂苷R0对人体QGY-7703肝癌细胞的增殖抑制作用。采用流式PI单染法,研究竹节香附素A、竹节香附皂苷R2及竹节香附皂苷R0对QGY-7703肿瘤细胞凋亡作用的影响。采用荧光分光光度计测定荧光强度的方法,研究不同结构两头尖皂苷单体对细胞内活性氧(ROS)含量的影响。结果:竹节香附素A、竹节香附皂苷R2、竹节香附皂苷R0及顺铂作用48小时后,对肝癌QGY-7703的IC50分别为5.11 g/ml,15.87 g/ml,28.39 g/ml,顺铂的IC50为5.27 g/ml。流式细胞仪检测结果显示,空白组G1期占百分数为36.19%、S期为59.11%、G2/M期为4.7%,竹节香附素A(5 g/ml)G1期为69.12%、S期为18.76%、G2期为12.12%;竹节香附皂苷R2(5 g/ml)G1期为60.60%、S期为30.35%、G2期为9.05%;竹节香附皂苷R0(5 g/ml)G1期为44.14%、S期为48.49%、G2期为7.37%;顺铂组(2.5 g/ml)G1期为47.49%、S期为42.71%、G2期为9.8%;荧光分光光度计测定DCF荧光强度结果为不同结构两头尖皂苷单体作用于QGY-7703细胞6h后,细胞内活性氧含量较空白组明显增加。结论:竹节香附素A、竹节香附皂苷R0及竹节香附皂苷R2可使QGY-7703细胞周期阻滞于G1期,并使细胞内产生过多的活性氧,从而加速诱导肿瘤细胞凋亡发挥抗癌作用,凋亡率随着侧链糖的个数增加而显著升高,并使细胞内产生过多的活性氧,从而加速诱导肿瘤细胞凋亡发挥抗癌作用。
OBJECTIVE: To study the antitumor activity of sarcandra amygdaloid A, Sarcandra glabra-root Saponins R2 and Saponins Saponins R0 in vitro, and to compare the effects of the number of side-chain sugars on antitumour activity. The content of reactive oxygen species in cells was studied in the process of apoptosis. Methods: The in vitro MTT assay was used to study the inhibitory effect of sarcaxin A, sarcandra saponin R2 and sarcandra ginsenoside R0 on the proliferation of human QGY-7703 hepatocarcinoma cells. The flow cytometry was used to study the effect of sarcaxin A, sarcandra saponin R2 and sarcandra ginsenoside R0 on the apoptosis of QGY-7703 tumor cells. Fluorescence spectrophotometer was used to measure the fluorescence intensity to study the effect of different structures of Saposaponins on the intracellular reactive oxygen species (ROS). Results: IC50 of QGY-7703 for hepatocarcinoma were 5.11 g / ml, 15.87 g / ml and 28.39, respectively after treated for 48 hours with flavonoids A, sylvite R2, slubbyosin R0 and cisplatin g / ml and cisplatin has an IC50 of 5.27 g / ml. Flow cytometry results showed that the percentage of G1 phase was 36.19% in the blank group, 59.11% in the S phase and 4.7% in the G2 / M phase. The Glutin A (5 g / ml) G1 phase was 69.12% The S phase was 18.76%, the G2 phase was 12.12%. The S phase was 60.60%, the S phase was 30.35%, the S phase was 9.05%. The Sjogrenoside R2 (5 g / ml) g / ml), G1 phase was 44.14%, S phase was 48.49%, G2 phase was 7.37%. In cisplatin group (2.5 g / ml), G1 phase was 47.49%, S phase was 42.71% and G2 phase was 9.8% DCF fluorescence intensity measured by the spectrophotometer results showed that the structure of reactive oxygen species in QGY-7703 cells treated with two different Saposaponins with different structures increased significantly compared with the blank group. Conclusion: Sarcandra fusiforme A, Slub Saponins R0 and Sarcandra saponin R2 can block the cell cycle of QGY-7703 in G1 phase and produce excessive reactive oxygen species in the cells, thus accelerating tumor induction Apoptosis play an anticancer role, the apoptosis rate increases significantly with the increase of the number of side chain sugars, and make the cells produce excessive reactive oxygen species, which can accelerate the apoptosis of tumor cells to exert anticancer effects.