加味逍遥散对肝癌细胞凋亡及钙离子的影响

Influence of Jiawei Xiaoyaosan on hepatoma cell apoptosis and calcium

目的观察加味逍遥散对离体肝癌SMMC-7721细胞生长的抑制作用及其对细胞内钙离子浓度的影响,探讨中药治疗肝癌的发病机制。方法采用MTT法测定高、中、低剂量加味逍遥散对肝癌SMMC-7721细胞生长的抑制作用;并利用流式细胞仪分析细胞凋亡情况;利用激光共聚焦显微镜观察细胞内游离钙离子情况。结果加味逍遥散对肝癌SMMC-7721细胞的生长有抑制作用,并且抑制率随着药物浓度的升高呈升高趋势;加味逍遥散高、中、低剂量组给药后凋亡率分别为0. 276、0. 202、0. 096;加入不同剂量的加味逍遥散时,钙离子荧光强度显示不同,低、中剂量时荧光增强不明显,而高剂量时细胞内荧光强度明显增强。结论加味逍遥散能抑制肿瘤细胞生长,促进细胞凋亡,同时能引起细胞内钙离子变化,这可能是加味逍遥散诱导肝癌SMMC-7721细胞凋亡的机制之一。

Objective To probe the inhibitory effect of human hepatocarcinoma cell line SMMC-7721 and the influence of intracellular free Ca2 +levels induced by Jiawei Xiaoyaosan,so as to reveal the pathogenesis of Chinese medicine treatment of liver cancer. Methods Determined by MTT method was used to determine the high,medium and low dose Jiawei Xiaoyaosan for liver cancer SMMC-7721 cell growth inhibition. The rate of apoptosis of liver cancer SMMC-7721 cell were examined by flow cytometry. The fluorescent intensity of intracellular free Ca2 +was observed by laser scanning confocal microscopy. Results Jiawei Xiaoyaosan had an inhibitory effect on the growth of liver cancer SMMC-7721 cells,and the inhibition rate increased with the increase of drug concentration. The apoptotic rate of Jiawei Xiaoyaosan was 0. 276 in the high,0. 202 in the medium and 0. 096 in the low dose groups.When adding different doses of Jiawei Xiaoyaosan,the fluorescence intensity of calcium ions was different. The fluorescence enhancement was not obvious at low and medium doses,while the intracellular fluorescence intensity was significantly enhanced at high doses. Conclusion Jiawei Xiaoyaosan can inhibit the proliferation of cancer cells and promote their apoptosis,and can cause changes in intracellular Ca2 +at the same time. This may be one of the mechanisms of Jiawei Xiaoyaosan to induce apoptosis of liver cancer SMMC-7721 cells.