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目的研究壁虎醇提物(GEE)对H22小鼠肿瘤组织血管新生的抑制作用及其机制。方法建立腋下移植H22肝癌模型,按照体重随机分为5组:模型组(0.9%Na Cl)、对照组(2 mg·kg~(-1)阿霉素)和高、中、低3个剂量实验组(GEE:80,40,20 mg·kg~(-1)),连续腹腔注射10 d,计算抑瘤率;用免疫组化法观察血管内皮细胞标记物(CD34)表达水平,计算新生微血管密度(MVD);以免疫印迹法检测血管内皮生长因子A(VEGF-A)、基质金属蛋白酶(MMP-2)、细胞外信号调节激酶(ERK)、磷酸化的细胞外信号调节激酶(P-ERK)的表达。结果高、中、低3个剂量实验组的抑瘤率分别为63.18%,50.91%,44.68%,说明GEE对H22小鼠移植瘤的抑瘤作用较明显。模型组小鼠肿瘤组织MVD为平均每个视野(24.7±4.1)个,与模型组比较,对照组、高、中、低3个剂量实验组的MVD分别为平均每个视野(13.6±3.6),(8.0±1.6),(11.0±1.1),(13.9±1.7)个,其中3个剂量实验组差异有统计学意义(均P<0.01)。模型组、对照组、低中高3个剂量实验组的VEGF-A与β-actin灰度比值平均分别为0.17,0.10,0.01,0.03,0.04;这5组的MMP-2与β-actin灰度比值平均分别为1.68,1.27,0.48,1.15,1.50,而P-ERK与ERK灰度比值平均分别为0.10,0.07,0.06,0.07,0.08,与模型组相比,这4组差异均有统计学意义(均P<0.01),说明GEE组和对照组的VEGF-A、MMP-2、P-ERK蛋白水平均明显减弱。结论 GEE可抑制小鼠H22肝癌移植瘤的生长,其抗肿瘤作用机制可能与降低肿瘤组织血管生成相关因子VEGF-A、MMP-2、P-ERK的表达及降低微血管密度有关。
Objective To study the inhibitory effect of gecko alcohol extract (GEE) on tumor angiogenesis in H22 mice and its mechanism. Methods The model of H22 hepatoma transplanted to the undectum was established and randomly divided into 5 groups according to body weight: model group (0.9% Na Cl), control group (2 mg · kg -1) doxorubicin), high, medium and low (GEE: 80, 40, and 20 mg · kg -1) were injected into the abdominal cavity for 10 days to calculate the tumor inhibition rate. The expression of CD34 was detected by immunohistochemistry (MVD). The expression of vascular endothelial growth factor A (VEGF-A), matrix metalloproteinase (MMP-2), extracellular signal regulated kinase (ERK), phosphorylated extracellular signal regulated kinase P-ERK) expression. Results The tumor inhibition rates of the high, middle and low doses of the three experimental groups were 63.18%, 50.91% and 44.68%, respectively, indicating that GEE has obvious anti-tumor effect on the H22 transplanted tumor. MVD in the model group mice was 24.7 ± 4.1 per field. Compared with the model group, MVD in the control, high, middle and low dose groups were respectively average (13.6 ± 3.6) , (8.0 ± 1.6), (11.0 ± 1.1) and (13.9 ± 1.7), respectively. There were significant differences in the three dose groups (all P <0.01). The average gray value of VEGF-A and β-actin in the model group, the control group, the low, middle and high dose groups were 0.17,0.10,0.01,0.03,0.04, respectively. The gray scale of MMP-2 and β-actin The average ratio of P-ERK to ERK was 0.10,0.07,0.06,0.07,0.08, respectively. Compared with the model group, the differences among the four groups were statistically significant (P <0.01), indicating that the protein levels of VEGF-A, MMP-2 and P-ERK in GEE group and control group were significantly decreased. Conclusions GEE can inhibit the growth of H22 hepatoma transplanted tumor in mice. The anti-tumor mechanism of GEE may be related to the decrease of the expression of VEGF-A, MMP-2, P-ERK and the decrease of microvessel density.