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目的:观察化痰祛瘀方含药血清对肝癌细胞血管内皮生长因子(VEGF)表达的影响。方法:健康SD大鼠15只,随机分为3组,大剂量组:化痰祛瘀方大剂量含药血清组,灌服化痰祛瘀方,给药剂量24g/kg(相当于临床等效剂量2倍);小剂量组:化痰祛瘀方小剂量含药血清组,灌服化痰祛瘀方,给药剂量12g/kg;正常血清组:灌服相应体积的生理盐水,共计5mL。通过MTT法检测化痰祛瘀方含药血清对肝癌细胞株增殖的抑制作用;通过Western bolt技术、免疫荧光技术观察VEGF蛋白表达的变化。结果:化痰祛瘀方含药血清能明显抑制人肝癌细胞株HCCLM3的增殖活力,不同剂量时(0g/kg、12g/kg、24g/kg)抑制率分别为1.60%、36.64%、63.92%,差异均有显著性意义(P<0.01);化痰祛瘀方含药血清能使肝癌细胞内的VEGF蛋白表达量下降,且随着化痰祛瘀方含药血清剂量增高(12g/kg、24g/kg),VEGF的蛋白表达呈下降趋势;免疫荧光试验显示,大剂量化痰祛瘀含药血清(24g/kg)可以完全抑制VEGF蛋白的表达。结论:化痰祛瘀方含药血清通过抑制VEGF的表达从而抑制肝癌细胞的生长。
Objective: To observe the effect of Huatanquyu Decoction-containing serum on the expression of vascular endothelial growth factor (VEGF) in hepatocellular carcinoma cells. Methods: Fifteen healthy SD rats were randomly divided into three groups. The high-dose group: high-dose phlegm and blood stasis group containing serum-containing group, gavage phlegm and blood stasis, the dose of 24g / kg (equivalent to clinical Low dose group: small dose of phlegm and blood stasis side containing serum group, gavage phlegm and blood stasis, the dose of 12g / kg; normal serum group: the corresponding volume of saline fed, the total 5mL. The inhibitory effect of Huatanquyu Decoction-containing serum on the proliferation of hepatoma cell line was detected by MTT assay. The expression of VEGF protein was observed by Western blot and immunofluorescence staining. Results: The serum of Huatanquyu prescription could significantly inhibit the proliferation of human hepatocellular carcinoma cell line HCCLM3. The inhibitory rates were 1.60%, 36.64% and 63.92% at different doses (0g / kg, 12g / kg and 24g / (P <0.01). The serum of Huatanquyu prescription could decrease the expression of VEGF protein in hepatocellular carcinoma cells. With the increase of the dose of Huatanquyu decoction (12g / kg , 24g / kg). The expression of VEGF protein showed a decreasing trend. Immunofluorescence assay showed that high dose of phlegm and blood stasis drug serum (24g / kg) could completely inhibit the expression of VEGF protein. Conclusion: The drug-containing serum of Huatanquyu prescription inhibits the growth of liver cancer cells by inhibiting the expression of VEGF.