【摘要】 目的:探讨活血化瘀方抑制小鼠Hepa1-6肝癌转移和生长的作用及其机制。方法:转染萤火虫荧光素酶基因(firefly luciferase gene)的C57BL/6小鼠肝癌Hepa1-6细胞,经筛选获得稳定表达GFP绿色荧光蛋白及荧光素酶基因的小鼠肝癌细胞株Hepa1-6 (luc2/GFP)。采用尾静脉注射及右侧腹部皮下注射的方法分别建立尾静脉肝癌转移模型及皮下肿瘤模型,并分别给予活血化瘀方52、104 g/kg组、索拉菲尼30 mg/kg组。造模后进行小动物成像荧光观测,16天后取皮下肿瘤组织,实时荧光定量PCR (q-PCR)法检测程序性死亡受体-1(PD-1)、白细胞介素-8 (IL-8)、肿瘤坏死因子-α(TNF-α)的表达;免疫组化(IHC)法检测CD4~+T细胞、CD8~+T细胞、血小板-内皮细胞粘附分子标记的微血管密度(CD31-MVD)。结果:活血化瘀方能抑制小鼠Hepa1-6肝癌的转移和生长,与模型组比较,活血化瘀方52 g/kg、104 g/kg治疗组可使PD-1、IL-8、TNF-αmRNA的表达明显下调;同时上调CD4~+T蛋白表达、升高CD4~+T/CD8~+T比值,下调CD8~+T、CD31MVD蛋白的表达。结论:活血化瘀方可能是通过调控CD4~+T细胞、CD8~+T细胞、PD-1、CD31、IL-8、TNF-α等多靶标来抑制小鼠Hepa1-6肝癌的转移和生长。更多还原

【Abstract】 Objective:To explore the effect of Huoxuehuayu formula on inhibiting the metastasis and growth of hepa1-6 liver cancer in mice and its mechanism. Methods:The C57 BL/6 cell line Hepa1-6, which was transfected with firefly luciferase gene, was used to obtain Hepa1-6(luc2/GFP) that expressing GFP green fluorescent protein and luciferase gene stably. Hepa1-6(luc2/GFP) cells were subcutaneously injected into the tail vein and the right abdomen of C57 mice to establish the model. Huoxuehuayu formula(52 g/kg,104 g/kg) and Sorafenib(30 mg/kg) were given respectively. IVIS Spectrum was performed. After 16 days,the subcutaneous tumor tissue was taken. CD4~+T cells, CD8~+T cells and CD31-MVD were detected by IHC, expressions of PD-1, IL-8 and TNF-α were detected by q-PCR.Results: Huoxuehuayu formula inhibited the metastasis and growth of hepa1-6 liver cancer in mice. Compared with the model group, expressions of PD-1, IL-8 and TNF-α were significantly decreased in 52 g/kg、104 g/kg Huoxuehuayu formula groups, the expressions of CD4~+T,CD4~+T/CD8~+T were up regulated, the expressions of CD8~+T and CD31-MVD were down regulated. Conclusion: Huoxuehuayu formula may inhibit the metastasis and growth of Hepa1-6 liver cancer in mice by regulating PD-1, IL-8, TNF-α,CD31, CD4~+T cells, CD8~+T cells.更多还原