miRNA-195抑制剂对高糖培养血管内皮细胞生物学行为的影响及机制

The effect of miRNA-195 inhibitor on the biological function of high-glucose cultured vascular endothelial cells and its mechanism

目的:探讨高糖环境下miRNA-195抑制剂对血管内皮细胞行为的影响及机制。方法:培养人脐静脉血管内皮细胞(HUVECs)ECV304,分别用高糖(HG)、miRNA-195模拟质粒(mimic)、miRNA-195抑制质粒(inhibitor)、Sirt1激动剂白藜芦醇(RES)处理细胞株,CCK-8试剂盒检测增殖活力,RT-PCR检测miRNA-195及Sirt1 mRNA水平,Western blot检测Sirt1、p-FoxO1、FoxO1、Caveolin-1、血管内皮生长因子(VEGF)蛋白表达水平,划痕实验检测HUVECs的迁移能力,transwell小室实验检测细胞侵袭能力,Matrigel胶检测HUVECs的成管能力。结果:HG促进HUVECs的增殖、迁移、侵袭,增强成管能力(P<0.05);而miRNA-195 inhibitor则明显抑制HUVECs的增殖、迁移,降低成管能力(P<0.05)。结论:miRNA-195 inhibitor或许通过miRNA-195/Sirt1/FoxO1/Caveolin-1/VEGF通路影响HG环境下HUVECs的生物学功能。

Objective:To explore the effect of miRNA-195 inhibitor on the biological function of vascular endothelial cells cultured by high glucose and its mechanism.Methods:Human umbilical vein endothelial cell line(HUVECs)ECV304 was culutred and treated with high glucose,miRNA-195 mimic,miRNA-195 inhibitor and Sirt1 agonist resveratrol.CCK-8 kit was used to detect proliferation activity,the levels of miRNA-195 and Sirt1 mRNA detected by RT-PCR,the protein expression levels of Sirt1,p-FoxO1,FoxO1,Caveolin-1,vascular endothelial growth factor(VEGF)detected by Western blot.The scratch test was used to detect the migration ability of cells,transwell chamber test was used to detect the invasion ability of cells.Matrigel glue was used to detect the ability of vascular endothelial cells to form tubes.Results:The proliferation,migration and tube formation of vascular endothelial cells were enhanced under high glucose conditions(P<0.05);while miRNA-195 inhibitor significantly inhibited the proliferation of vascular endothelial cells and reduced the ability of tube formation(P<0.05).Conclusion:MiRNA-195 inhibitor may affect the biological function of endothelial cells through miRNA-195/Sirt1/FoxO1/Caveolin-1/VEGF pathway under high glucose conditions.