健脾补肾解毒方调节HIF⁃1α对裸鼠肝癌血管新生的影响

Influence of Jianpi Bushen Jiedu Recipe on angiogenesis of liver cancer in nude mice by regulating HIF⁃1α

目的:评价健脾补肾解毒方通过缺氧诱导因子1α(HIF-1α)对裸鼠肝癌移植瘤及血管新生相关因子的抑制作用。方法:将接种肝癌皮下移植瘤的裸鼠随机分为空白组、索拉非尼组及健脾补肾解毒方高、中、低剂量组,分别给予相应药物干预,连续4周。比较各组裸鼠移植瘤大小;通过免疫组化法检测各组肿瘤微血管密度(MVD),采用荧光Real-time PCR法检测HIF⁃1αmRNA,酶联免疫吸附(ELISA)法检测不同药物干预下对移植瘤一氧化氮合酶(NOS)、血管内皮生长因子(VEGF)及碱性成纤维细胞生长因子(bFGF)的影响。结果:与空白组相比,健脾补肾解毒方各剂量组瘤体均明显减小(P<0.05),MVD值均明显降低(P<0.01),索拉非尼组MVD值下降(P<0.05);与空白组相比,健脾补肾解毒方高剂量组、索拉非尼组HIF-1αmRNA表达均明显减少(P<0.05);与空白组相比,健脾补肾解毒方高剂量组、索拉非尼组VEGF表达均明显下调(P<0.05),且该两组bFGF表达较空白组也均明显减少(P<0.05)。结论:健脾补肾解毒方可以缩小裸鼠肝癌移植瘤,抑制移植瘤血管生成。其作用机制可能是通过下调HCCLM3细胞HIF-1α的表达,减少VEGF和bFGF的表达,同时降低移植瘤NOS活性,从而抑制肝癌血管新生。

Objective:To evaluate the inhibitory effects of Jianpi Bushen Jiedu Recipe(JBJR)on hepatocellular xenograft tumor and angiogenesis-related factors in nude mice through hypoxia-inducible factor-1α(HIF-1α).Methods:The nude mice vaccinated with subcutaneous transplantation tumor of liver cancer were randomly divided into the blank group,sorafenib group,and JBJR high,medium and low dose groups.The mice were given corresponding drug intervention for four consecutive weeks.The transplantation tumor size was compared among the groups.The microvessel density(MVD)of tumor was detected by immunohistochemistry,the HIF-1αmRNA was detected by real-time PCR,and the influences of different drugs on nitric oxide synthase(NOS),vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)were detected by enzyme linked immunosorbent assay(ELISA).Results:Compared with the blank group,the tumor volume in each dose group of JBJR was significantly reduceed(P<0.05),the MVD were significantly decreased(P<0.01),and the MVD of the sorafenib group was decreased(P<0.05).Compared with the blank group,the HIF-1αmRNA expression in the high dose group of JBJR and sorafenib group were significantly reduced(P<0.05).Compared with the blank group,the expression of VEGF in the high dose group of JBJR and sorafenib group was significantly down-regulated(P<0.05),and the expression of bFGF in above two groups were also significantly reduced compared with the blank group(P<0.05).Conclusion:JBJR can reduce the transplantation tumor of liver cancer in nude mice,and inhibit the angiogenesis of transplantation tumor.Its mechanisms may be reducing the expressions of VEGF and bFGF,meanwhile decreasing the activity of NOS in transplantation tumor,thereby inhibiting angiogenesis of liver cancer through down-regulating the expression of HIF-1αin HCCLM3 cells.