落新妇苷对人脐静脉内皮细胞迁移及管腔形成的影响

Effects of Astilbin on the migration and angiogenesis of vascular endothelial cells

摘要:

目的 探讨落新妇苷在体外促进血管生成过程中的影响。 方法 将体外培养的人脐静脉内皮细胞 （HUVECs）中分别加入不同浓度（0、1、10、20、40 µmol/L）的落新妇苷，通过 MTT检测HUVECs的增殖能力；划痕实 验检测细胞迁移能力；基质胶小管形成实验检测细胞管腔形成能力；Western blot检测血管内皮生长因子A（VEGFA）、 血管内皮生长因子受体 2（VEGFR-2）及血管性血友病因子（VWF）的蛋白表达情况。结果 将不同浓度落新妇苷处理 HUVECs 6、12 和 24 h 后，细胞的增殖数目与对照组相比差异无统计学意义（P>0.05），而落新妇苷作用细胞 48 h后，40 µmol/L组细胞的增殖数与各组相比差异均有统计学意义（P<0.01 或 0.001）。划痕实验显示 10 µmol/L 组 6、12 和 24 h 有 大量的细胞向中间的划痕区域迁移，细胞分布广泛，较对照组显著增多（P<0.01）；24 h时，10 µmol/L 组划痕愈合面明显 多于 1、20 µmol/L 组（P<0.01）。与对照组比较，10 µmol/L组形成的管腔样结构最多，各浓度落新妇苷处理组形成的节 点数、主节段长度及网格数均显著增加（P<0.05）。落新妇苷能显著增加VEGFA、VEGFR-2 和VWF的蛋白表达，1、10 µmol/L组较对照组明显增高（P<0.01）。结论 落新妇苷可促进HUVECs细胞迁移及管腔形成。

Abstract:

Objective The aim of this study is to investigate the effect of astilbin on angiogenesis in vitro. Methods Different concentrations (0, 1, 10, 20, 40 μmol/L) of astilbin were added into HUVECs cultured in vitro, and the proliferation ability of HUVECs was detected by using MTT. The ability of cell migration was detected by scratch test, and the ability of cell lumen formation was determined by using matrix glue tubule formation test. The expression of angiogenesis-related factors (VEGFA, VEGFR-2, and VWF) was detected by using Western blot. Results There was no significant difference in the number of HUVECs in astilbin treatment with different concentrations compared with the control group (P>0.05), while the number of cells in the 40 μmol/L group was statistically different from that of the control group after 48 h (P<0.01 or 0.001). The Scratch test showed that a large number of cells migrated to the middle scratch area in the 10 µ mol/L group at 6, 12 and 24 h, with a wide distribution of cells, significantly increasing compared to the control group, and the difference was statistically significant (P<0.01). The migration of HUVECs in the 10 µmol/L group was significantly increased compared to the 1 µmol/ L group and the 20 µmol/L group at 24 h (P<0.05). Compared with the control group, the most lumen-like structures were formed in the 10 µmol/L group, and the number of nodes, main segment length and mesh number were significantly increased in the astilbin treatment groups at all concentrations (P<0.05). Astilbin could significantly increase the expression of angiogenic factors VEGFA, VEGFR-2 and VWF, which were significantly higher in 1 and 10 μmol/L groups than in the control group (P<0.01). Conclusion Astilbin can promote the migration and angiogenesis of HUVECs cells.