杠柳苷对H_(22)荷瘤小鼠的抑瘤作用及其机制研究

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目的研究香加皮中杠柳苷体内抗肿瘤作用,探讨其可能作用机制。方法将小鼠肝癌H22细胞经体外培养并于BALB/c小鼠腹腔内传代后,无菌接种于小鼠皮下,建立荷瘤小鼠模型。采取不同剂量杠柳苷(0.25、0.50、1.00 mg/kg)ip给药,每天1次,连续15 d,观察荷瘤小鼠肿瘤生长情况,测定抑瘤率,应用流式细胞术检测杠柳苷对肿瘤细胞周期与凋亡的影响;采用透射电镜观察荷瘤小鼠肿瘤细胞超微结构的变化。结果杠柳苷对小鼠H22皮下移植瘤具有显著的抑制作用,小鼠成瘤时间明显晚于模型组,肿瘤生长缓慢,低、中、高3个剂量组抑瘤率分别为60.28%、68.09%、74.74%。流式细胞术分析结果显示,不同剂量杠柳苷处理小鼠移植瘤细胞后,处于G0/G1期细胞明显增多,S期和G2/M期细胞显著减少,与模型组相比差异显著(P<0.05),其中1.00 mg/kg杠柳苷组G0/G1期细胞由对照组的(46.90±5.80)%升高至(83.80±2.52)%,S期和G2/M期细胞分别由对照组的(38.30±5.11)%、(14.80±0.70)%下降至(5.33±2.73)%、(10.87±0.25)%;而且杠柳苷各组小鼠肿瘤细胞的凋亡率均明显增加,其中1.00 mg/kg组凋亡率高达(32.35±1.75)%,并可见典型的凋亡峰。透射电镜观察可见,杠柳苷各组小鼠肿瘤组织中出现大量典型的凋亡细胞,而对照组瘤组织内多为富含线粒体与内质网的代谢旺盛的肿瘤细胞。结论杠柳苷具有很强的体内抗肿瘤作用,其作用机制可能与阻滞细胞周期和诱导肿瘤细胞凋亡有关。 Objective To study the antitumor effect of periadalcins in Xiangkapi and to explore its possible mechanism. Methods Mouse hepatocellular carcinoma H22 cells were cultured in vitro and passaged in BALB / c mice intraperitoneally. Then the cells were aseptically subcutaneously inoculated into mice to establish a tumor-bearing mouse model. Take different doses of bariposide (0.25,0.50,1.00 mg / kg) ip administration, once a day for 15 days, observe tumor growth in tumor-bearing mice, determination of tumor inhibition rate, the application of flow cytometry to detect Glycosides on cell cycle and apoptosis of tumor cells. The ultrastructure of tumor cells in tumor-bearing mice was observed by transmission electron microscope. Results Periplocin had a significant inhibitory effect on subcutaneously transplanted H22 mice. The tumor formation time of the mice was significantly later than that of the model group, and the tumor growth was slow. The tumor inhibition rates of low, medium and high dose groups were 60.28% and 68.09 %, 74.74%. The results of flow cytometry showed that the cells in G0 / G1 phase were significantly increased, and the cells in S phase and G2 / M phase were significantly decreased compared with the model group (P (P <0.05). The cells in G0 / G1 phase at the concentration of 1.00 mg / kg were increased from (46.90 ± 5.80)% to (83.80 ± 2.52)% in control group and the cells in S phase and G2 / (38.30 ± 5.11)% and (14.80 ± 0.70)%, respectively, to (5.33 ± 2.73)% and (10.87 ± 0.25)%, respectively. Moreover, the apoptosis rate of tumor cells in all groups increased significantly The apoptosis rate in mg / kg group was as high as (32.35 ± 1.75)%, and the typical apoptotic peak was seen. Transmission electron microscopy showed that a large number of typical apoptotic cells were found in the tumor tissues of mice in each group, whereas in the control group, most of them were metabolically rich tumor cells rich in mitochondria and endoplasmic reticulum. Conclusion Periplocin has a strong anti-tumor effect in vivo, and its mechanism may be related to arresting cell cycle and inducing tumor cell apoptosis.
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