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褐藻糖胶对RPMI 8226细胞血管生成的影响

ISSN号：1000-4718
期刊名称：《中国病理生理杂志》
时间：0
分类：R363[医药卫生—病理学;医药卫生—基础医学] R979.1[医药卫生—药品;医药卫生—药学]
作者机构：[1]重庆医科大学附属第一医院血液科,重庆400016, [2]重庆医科大学附属第一医院中心实验室,重庆400016, [3]重庆市三峡中心医院血液科,重庆404000
相关基金：重庆市卫生局重点项目（No.2013-1-013）

关键词：褐藻糖胶, 多发性骨髓瘤, HIF-1Α, VEGF, AKT, ERK1/2, Fucoidan, Multiple myeloma, HIF-1α, VEGF, AKT, ERK1/2

中文摘要：

目的：探讨褐藻糖胶对多发性骨髓瘤RPMI 8226细胞血管生成的影响及可能的作用机制。方法：体外培养多发性骨髓瘤RPMI 8226细胞及人血管内皮细胞EA.hy 926细胞,采用MTT法检测褐藻糖胶对RPMI8226细胞活力的影响;流式细胞术检测细胞周期及凋亡率;ELISA法检测褐藻糖胶处理RPMI 8226细胞后的培养液上清中VEGF的含量;小管形成实验观察褐藻糖胶处理RPMI 8226细胞后的培养液上清对EA.hy 926细胞诱导小管形成能力的影响;Western blot检测HIF-1α、VEGF、p-AKT和p-ERK1/2的蛋白水平。结果：褐藻糖胶对RPMI8226细胞活力的抑制作用呈浓度及时间依赖性;褐藻糖胶作用RPMI 8226细胞72 h后,细胞周期被阻滞于G1期,细胞凋亡率呈浓度依赖性明显增高,各组均高于对照组（P〈0.05）;ELISA法结果显示褐藻糖胶处理72 h后的细胞培养液上清中VEGF的含量明显减少,与对照组相比,处理组上清中VEGF的含量下降（P〈0.05）;内皮细胞形成小管数目与面积随着褐藻糖胶的浓度升高而减小,100 mg/L组差异显著（P〈0.05）;Western blot结果显示HIF-1α、VEGF、p-AKT和p-ERK1/2的蛋白水平与褐藻糖胶浓度呈负相关（P〈0.05）。结论：褐藻糖胶减少骨髓瘤细胞自分泌VEGF,减弱血管内皮细胞形成小管的能力,降低HIF-1α和VEGF的蛋白水平,这可能与抑制AKT和ERK1/2的磷酸化有关。

英文摘要：

AIM： To investigate the effects of fucoidan on the angiogenesis of multiple myeloma cells in vitro,and its related mechanisms. METHODS： The human multiple myeloma RPMI 8226 cells and human endothelial cells were cultured in vitro. The growth inhibition rate of RPMI 8226 cells was examined by MTT assay. The cell cycle and apoptosis rate were measured by flow cytometry. RPMI 8226 cells were treated with fucoidan for 72 h,and the cell culture supernatant was collected. The VEGF concentration was examined by ELISA,and the tube formation assay was applied to assess the angiogenic activity. After treatment with fucoidan for 72 h at different concentrations,the protein levels of HIF-1α,VEGF,p-AKT and p-ERK1 /2 were detected by Western blot. RESULTS： Fucoidan inhibited the growth of RPMI 8226 cells in a dose- and time-dependent manner. After treatment with fucoidan for 72 h,the cell cycle was arrested at G1 phase,and the apoptotic rate of RPMI 8226 cells was increased with the increasing concentration of fucoidan,which was much higher than that in control group（ P〈0. 05）. The VEGF concentration was significantly decreased with the increasing concentration of fucoidan. The numbers and areas of the capillary-like structures decreased while the concentration of fucoidan increased,and those at 100 mg / L were less than those in the control（ P〈0. 05）. The protein levels of HIF-1α,VEGF,p-AKT and p-ERK1 /2 in fucoidan group were significantly lower than those in control group（ P〈0. 05）. CONCLUSION： Fucoidan inhibits the secretion of VEGF in multiple myeloma cells,and reduces angiogenesis induced by multiple myeloma cells. It inhibits the protein expression of HIF-1α and VEGF,which may be related to inhibiting the phosphorylation of AKT and ERK1 /2.

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