地锦草调控NF-κB/VEGF信号通路抑制肿瘤血管生成的机制

Mechanism of Humifuse euphorbia in Regulating Tumor Angiogenesis by NF-κB/VEGF Signal Pathway

目的：通过观察地锦草对H22肝癌小鼠核转录因子-κB/血管内皮生长因子（nuclear factor kappa-B/vascular endothelial growth factor,NF-κB/VEGF）信号通路相关蛋白的影响,探讨地锦草抑制肿瘤血管生成的机制。方法：昆明种小鼠皮下接种肝癌H22细胞建立H22肝癌小鼠模型,随机分为模型组,环磷酰胺组,地锦草组高、中、低剂量组。地锦草低、中、高剂量组分别灌胃（ig）给药1.13,2.25,4.50 g·kg^-1,环磷酰胺组腹腔注射（ip）给药50 mg·kg^-1,模型组和环磷酰胺组灌服相同体积的蒸馏水,14 d后处死小鼠,测量肿瘤质量和小鼠体质量,计算抑瘤率;免疫组织化学检测微血管密度标记物血小板-内皮细胞黏附分子31（platelet endothelial cell adhesion molecule-31,CD31）,聚合酶链反应检测肿瘤组织NF-κB,肿瘤坏死因子（TNF）-α,VEGF mRNA表达,免疫组化检测NF-κB,TNF-α,VEGF蛋白表达。结果：与模型组比较,给予地锦草后,H22肝癌小鼠瘤重减轻（P〈0.01）,地锦草高剂量组抑瘤率升高,高达59.1%（P〈0.01）;肿瘤组织CD31表达明显下降（P〈0.01）;肿瘤组织NF-κB,TNF-α,VEGF mRNA和蛋白表达均下降（P〈0.01）。结论：地锦草对H22肝癌小鼠肿瘤的抑制作用可能是通过减少肿瘤新生血管的生成,与减弱肿瘤组织的炎性因子表达有关。

Objective： To observe the mechanism of Humifuse euphorbia on tumor angiogenesis in H22 liver cancer mice by regulating signal pathway of nuclear factor kappa-B/vascular endothelial growth factor（ NF-κB/VEGF）. Method： The models of SPF grade Kunming mice with H22 liver cancer were established through armpit injection. The animals with successful modeling were randomly divided into five groups： model control group,cyclophosphamide group（ ip,50 mg·kg^-1）,high-dose group（ ig,4. 50 g·kg^-1）,middle-dose group（ ig,2. 25 g·kg^-1） and low-dose group（ ig,1. 13 g·kg^-1）,with 10 mice in each group. Their weights and tumor weights were measured after the 14-day drug administration. Index of tumor was calculated. Tumor microvessel density, which was marked as platelet endothelial cell adhesion molecule-31, was detected by immunohistochemitry. Tumor tissue NF-κB,VEGF,tumor necrosis factor-α（ TNF-α） mRNA expressions were detected by polymerase chain reaction and protein expression by immunohistochemitry. Result： Compared with model group,H. euphorbia could significantly reduce tumor weigh（ P〈0. 01）. High-dose group showed an inhibition rate of 59. 1% for H22 liver cancer in mice. CD31 expression of the high-dose group was significantly lower than that of the model group,while the expressions of NF-κB,TNF-α,VEGF mRNA and protein in tumor tissue were significantly decreased（ P〈0. 01）. Conclusion： H. euphorbia can inhibit the growth of H22 liver cancer mice,and its mechanism is probably related to the inhibition of the growth of tumor angiogenesis,which may activate NF-κB/VEGF signaling pathway.