加味逍遥散对肝癌细胞凋亡及钙离子的影响
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摘要
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.
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.
引文
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[10]朱沛维.六味地黄汤合丹栀逍遥散加减改善乳腺癌三苯氧胺不良反应患者的效果分析[J].临床合理用药,2017,10(9C):15-6.
[11]郭维,孙琪,桂牧微,等.逍遥散对慢性应激大鼠模型血清肿瘤标志物的影响[J].时珍国医国药,2017,28(4):807-9.
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[2] Torre L A,Bray F,Siegel R L,et al. Global cancer statistics,2012[J]. CA Cancer J Clin,2015,65:87-108.
[3] Chen W,Zheng R,Baade P D,et al. Cancer statistics in China,2015[J]. Can J Clin,2016,66(2):115-32.
[4]占义平,凌昌全.原发性肝癌中医证候研究概述[J].中医杂志,2017,58(2):167-70.
[5]王磊,周荣耀.周荣耀补肾健健脾法治疗原发性肝癌经验[J].辽宁中医杂志,2014,41(2):2547-8.
[6]宋慧娴,乔飞,邵铭.中医药治疗原发性肝癌的研究进展[J].临床肝胆病杂志,2016,32(1):174-7.
[7]史国军,叶兴涛,陆宁,等.键脾柔肝方联合替吉奥胶囊治疗中晚期原发性肝癌患者30例临床观察[J].中医杂志,2016,57(8):677-81.
[8]山广志,邱慧颖,史国军,等.健脾柔肝汤对荷肝癌H22小鼠肿瘤组织VEGF表达的影响[J].中华中医药学刊,2014,32(10):2465-8.
[9]黄静,马萍,卢建峰.中医药干预肝癌干细胞的研究进展[J].世界最新医学信息文摘,2018,18(43):74-6,78.
[10]朱沛维.六味地黄汤合丹栀逍遥散加减改善乳腺癌三苯氧胺不良反应患者的效果分析[J].临床合理用药,2017,10(9C):15-6.
[11]郭维,孙琪,桂牧微,等.逍遥散对慢性应激大鼠模型血清肿瘤标志物的影响[J].时珍国医国药,2017,28(4):807-9.
[12]朱政,杨昌卫,陈永强,等.丹栀逍遥散联合消融治疗肝脏肿瘤的临床观察[J].中医临床研究,2017,9(8):54-5.