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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: Clover green has a good clinical basis for anti-hepatocellular carcinoma, and the extract of total flavonoids from Radix Tetrastigmae (TF) has particularly significant anticancer activity. This study from cell-animal experiments clear clover total flavonoids anti-liver cancer, clover leaf for the treatment of liver cancer to establish the material basis of efficacy. Methods: The CCK-8 method was used to detect the inhibitory effect of total flavonoids of clover leaf on the proliferation of hepatocellular carcinoma cells. The IC50 of the drug was calculated and the high, medium and low doses of TF were determined. The HepG2 cells were stained with Annexin V-FITC / PI double staining and the apoptosis rate of HepG2 cells was detected by flow cytometry. The establishment of human hepatoma cell line HepG2 xenografts in nude mice model to observe the TF of HepG2 xenografts in nude mice body weight, tumor growth volume, inhibition rate. Results: TFIC50 was 3.247 mg / m L, TF high, medium and low doses were 5,1.25,0.312 5 mg / m L; compared with the control group, high, medium and low doses of TF 48 h after the treatment of liver cancer HepG2 cells were significantly inhibited (P <0.01). At 24 h after TF treatment, the apoptosis of HepG2 cells was significantly increased And gradually increased with the increase of concentration, all of which had significant difference (P <0.01). The inhibition rates of high, medium and low doses of TF were 64.07%, 53.64% and 46.69%, respectively. The high dose of TF inhibited tumor growth of hepatoma mice better than that of CTX (cyclophosphamide) The inhibitory effect on tumor growth in HCC mice was weaker than CTX. CONCLUSION: Total flavonoids (high, medium and low concentrations) of Clovervehyl can obviously inhibit the proliferation of HepG2 cells and increase gradually with the increase of concentration. With the increase of concentration gradually increased. High dose of TF (15g / kg) was more effective than CTX in inhibiting tumor growth in HCC mice, while the low dose of TF (7.5,3.75g / kg) was less effective than CTX in tumor growth of HCC mice.