L-岩藻糖对大鼠肝癌射频消融术后乏氧微环境中HIF-1α、MMP-2、VEGF表达的影响

The research of expression of HIF-1α、MMP-2、VEGF in hypoxia environmental after radiofrequency ablation of rat hepatocellular carcinoma model treated with L-fucose

目的:观察L-岩藻糖对大鼠肝癌射频消融术后乏氧微环境中乏氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP-2)表达的影响。方法:采用大鼠肝叶内瘤块种植法建立大鼠种植性肝癌模型,建模成功后随机分为5组,非射频消融组、射频消融+生理盐水对照组、射频消融+L-岩藻糖大剂量组、射频消融+L-岩藻糖中剂量组、射频消融+L-岩藻糖小剂量组。对于非射频消融组大鼠只进行穿刺操作,不进行射频消融术,其余4组大鼠均进行射频消融术,术后分别给予不同剂量L-岩藻糖或生理盐水灌胃。术后21天后处死大鼠,取出瘤块,采用免疫组化法检测瘤块中HIF-1α、VEGF及MMP-2的表达情况。结果:射频消融+生理盐水对照组大鼠肿瘤周边组织中HIF-1α表达明显增高,而射频消融+岩藻糖大剂量组大鼠其表达显著降低;3岩藻糖组大鼠瘤周组织中MMP-2表达均降低,其中岩藻糖小剂量组降低最为显著;VEGF阳性染色主要集中在肿瘤细胞膜和细胞质,3岩藻糖组大鼠VEGF表达减少,VEGF在3个剂量组间的表达比较差异有明显意义。结论:肝癌大鼠射频消融术后HIF-1α显著增高,而L-岩藻糖能明显降低射频消融术后残留肿瘤局部尤其是癌旁组织中的HIF-1α表达水平,显著降低肿瘤局部微环境中MMP-2合成,残留肿瘤及癌旁组织中的HIF-1α与MMP-2的表达呈相关性;3岩藻糖组大鼠VEGF表达降低,组间比较差异显著,提示具有一定的剂量依赖性。

Objective:To observe the effect of L-fucose on the expression of HIF-1an of rat hepatocellular carcinoma model.Methods:Rat hepatocellular carcinoma model was established by implanting Walker-256 tumor tissue blocks under hepatic capsule.The models were successfully established and randomly divided into five groups:non-radiofrequency ablation group,radiofrequency ablation+NS control group,radiofrequency ablation+L-fucose high-dose group,radiofrequency ablation+L-fucose medium-dose group,radiofrequency ablation+L-fucose low-dose group.Rats in the non-radiofrequency ablation group received only puncture without radiofrequency ablation,while rats in the other four groups underwent radiofrequency ablation and were given different doses of L-fucose intragastrically after operation.After 20 days,rats were sacrificed and the tumors were removed.The expressions of HIF-1a,VEGF and MMP-2 in the tumors were detected by immunohistochemistry.Results:The expression of HIF-1alpha in the peritumoral tissues of the radiofrequency ablation+saline group was significantly increased,while that in the radiofrequency ablation+high dose group was significantly decreased.Compared with the radiofrequency ablation+saline group,the expression of MMP-2 in the peritumoral tissues of the three dose groups of fucoida decreased,especially in the low dose group.The positive staining of vascular endothelial growth factor mainly concentrated in the tumor cell membrane and cytoplasm.The expression of vascular endothelial growth factor decreased in all three dose groups,and there was a significant difference among the three dose groups,suggesting a dose-dependent manner.Conclusion:The expression of HIF-1alpha increased after operation,and L-fucose significantly decreased the expression of hypoxia-inducible factor-1alpha in local tumors,especially around tumors.L-fucose significantly decreased the synthesis of MMP-2 in local microenvironment of tumors.There was a positive correlation between the expression of HIF-1alpha and MMP-2 in hepatocellular carcinoma.The expression of vascular endothelial growth factor decreased in all three dose groups,and there was a significant difference in the expression of vascular endothelial growth factor between the three dose groups,suggesting a dose-dependent manner.