三叶青总黄酮对人肝癌HepG-2细胞及裸鼠异种移植瘤的药效作用研究  

Study on Effect of Total Flavonoids from Radix Tetrastigmae on Human Hepatocellular Carcinoma HepG-2 Cells and Xenograft Tumor in Nude Mice

作　　者：汪正飞[1] 杨祝仁 张轶雯[3] 李清林[3] 赵岩峰[4] 

机构地区：[1]衢州市人民医院肝胆外科,浙江衢州324000 [2]辽宁省药品认证中心,辽宁沈阳110036 [3]浙江省肿瘤医院,浙江杭州310022 [4]北京中医药大学东方学院,北京100029

出　　处：《中华中医药学刊》2017年第10期2615-2620,I0016,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81503165);浙江省中医药科技计划项目(2015ZQ008)

摘　　要：目的:三叶青抗肝癌作用具有良好的临床基础,而三叶青总黄酮(Total flavonoids from Radix Tetrastigmae,TF)提取物的抗癌活性尤其显著。本研究从细胞-动物实验明确三叶青总黄酮抗肝癌作用,为三叶青治疗肝癌建立药效物质基础。方法:通过CCK-8法检测三叶青总黄酮对肝癌细胞增殖的抑制作用,并计算药物的IC50,确定TF的高、中、低剂量。采用Annexin V-FITC/PI双染法染色HepG2细胞,采用流式细胞仪检测HepG2细胞凋亡率。通过建立人肝癌细胞系HepG2移植瘤裸鼠模型,观察TF对HepG2移植瘤裸鼠模型的体质量、肿瘤生长体积、抑瘤率的影响。结果:TFIC50为3.247 mg/m L,TF的高、中、低的剂量分别为5、1.25、0.312 5 mg/m L;与对照组比较,TF高、中、低浓度剂量48 h后对肝癌HepG2细胞增殖均有明显的抑制作用,且随浓度的增加而逐渐增强,均具有显著差异(P<0.01);TF高、中、低浓度剂量24 h后均有明显促进肝癌HepG2细胞的凋亡作用,且随浓度的增加而逐渐增强,均具有显著差异(P<0.01)。TF的高、中、低剂量的抑瘤率分别为64.07%、53.64%、46.69%,TF高剂量对肝癌小鼠肿瘤生长的抑制作用优于CTX(环磷酰胺),而TF中、低剂量对肝癌小鼠肿瘤生长的抑制作用弱于CTX。结论:三叶青总黄酮(高、中、低浓度)对肝癌HepG2细胞增殖均有明显的抑制作用,且随浓度的增加而逐渐增强;对肝癌细胞的凋亡均有明显促进的作用,且随浓度的增加而逐渐增强。TF高剂量(15g/kg)对肝癌小鼠肿瘤生长的抑制作用优于CTX,而TF中、低剂量(7.5、3.75 g/kg)对肝癌小鼠肿瘤生长的抑制作用弱于CTX。Objective： To investigate the antitumor activity of total flavonoids from Radix Tetrastigmae（TF） which has a good clinical basis. In this study, the effects TF on liver cancer were determined from cell and animal experiments. Methods ： CCK - 8 method was used to detect the inhibitory effect of TF on the proliferation of human hepatocellular carcinoma ceils and the ICs0 was calculated to determine the high, middle and low doses of TF. HepG2 ceils were stained with Annexin double staining method. The apoptosis rate of HepG2 cells was detected by flow cytometry （ Annexin V - FITC/ PI）. The nude mice model of human hepatocellular carcinoma cell line HepG2 was established to observe the effect of TF on the body weight, tumor growth volume and tumor inhibition rate of HepG2 transplanted tumor nude mice model. Results： When TF IC50 was 3. 247mg/mL, high, medium and low doses of TF were 5 mg/mL, 1.25 mg/mL and 0. 312 5 mg/mL. Compared with the control group, TF high, medium and low doses of 48 h on the proliferation of hepatocellular carcinoma HepG2 cells was significantly inhibited and with the concentration increased, there were significant differences （P〈0. 01）. TF high, medium and low doses of 24 h can obviously promote the apoptosis of hepatocellular carcinoma cell line HepG2, and with the concentration increased, there were significant differences （P 〈 0. 01 ）. The inhibition rates of TF high, medium and low doses were 64.07%, 53.64% and 46.69%, better than the CTX inhibitory effect of high dose of TF on tumor growth in mice and the inhibitory effect （eyclophosphamide）. The low dose TF on tumor growth in mice was weaker than CTX. Conclusion： TF（high, middle and low concentrations） inhibited proliferation of HepG2 cells and also increased with the increasing concentration. The effect of apoptosis on hepatoma cells was significantly promoted with the concentration increased. TF high dose （15 g/kg） was more effective than CTX in inhibiting the growth of liver cancer in mice, and

关 键 词：肝癌 三叶青 TF HEPG-2细胞 药效 

分 类 号：R285.5[医药卫生—中药学]