基于毒痰瘀虚理论探讨中药复方对肝癌血管生成的影响

The Effects of Herbal Compound on Hepatocellular Carcinoma Angiogenesis Based on the Theory of Toxin/Phlegm/Stagnation-induced Deficiency

目的:基于毒痰瘀虚理论探讨中药复方对肝癌血管生成的影响。方法:制备Wistar大鼠含药血清,以含药血清体外培养HepG2细胞,并分为空白组、中药复方组(毒痰瘀脾肾虚方、毒痰瘀脾虚方、毒痰瘀肾虚方、毒痰瘀肾阳虚方、毒痰瘀肾阴虚方、毒痰瘀方);采用酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)法检测细胞培养上清液中血管内皮生长因子(vascular endothelial growth factor,VEGF)表达;采用HepG2肝癌移植性肿瘤模型方法于小鼠腋下接种HepG2肝癌细胞株,免疫组织化学法检测肿瘤微血管密度(MVD)。结果:与空白组比较,中药复方组可降低肝癌HepG2细胞中VEGF表达(P<0.01),其中毒痰瘀脾肾虚方(全方)组最明显。而除全方组外,其余各中药复方组对肝癌HepG2细胞中VEGF表达的影响作用相当,差异无统计学意义(P>0.05)。与空白组相比,中药复方组均能降低肝癌HepG2小鼠肿瘤组织中MVD,毒痰瘀脾肾虚方组最明显,差异具有统计学意义(P<0.05);毒痰瘀组与毒痰瘀脾肾虚方组相比差异有统计学意义(P<0.05)。结论:基于毒痰瘀虚理论组方的中药复方具有抑制肝癌血管生成的作用。

Objective:To discuss the influence of herbal compound on hepatocellular carcinoma(HCC)angiogenesis on the foundation of the theory of deficiency induced by toxin,phlegm and stagnation.Methods:After preparing drug-containing serum from Wistar rats,HepG2 cells were cultured in vitro with drug-containing serum,divided into the blank group and herbal compound groups[toxin/phlegm/stagnation-induced spleen-kidney deficiency prescription(SKD),toxin/phlegm/stagnation-induced spleen deficiency prescription(SD),toxin/phlegm/stagnation-induced kidney deficiency prescription(KD),toxin/phlegm/stagnation-induced kidney Yang deficiency prescription(KYAD),toxin/phlegm/stagnation-induced kidney Yin deficiency prescription(KYID),toxin,phlegm,stagnation prescription(TPS)];ELISA method was adopted to detect the expressions of VEGF in the cell liquid supernatant;immunohistochemical method was utilized to measure tumor microvessel density(MVD)by using HepG2 hepatoma xenograft tumor model via the axillary inoculation of HepG2 hepatoma cell line.Results:Compared with the blank group,herbal compound groups could lower the expressions of VEGF in HepG2 cells(P<0.01),among them,the groups of whole prescription showed the obvious effects.However,except the group of SKD(whole prescription),the rest of herbal compound groups showed equivalent effects on,SD the expressions of VEGF in HepG2 cells,and the difference had no statistical meaning(P>0.05).Compared with the blank group,herbal compound groups could reduce MVD in tumor tissue of HCC HepG2 mice,SKD presenting the most obvious effects,and the difference had statistical meaning(P<0.05);the difference had statistical meaning when TPS was compared with SKD(P>0.05).Conclusion:Herbal compound formed by the theory of toxin/phlegm/stagnation-induced deficiency could inhibit HCC angiogenesis.