Silva Carolina, Rodrigues Ilda, Andrade Sara, Costa Raquel, Soares Raquel
Department of Biomedicine, Unit of Biochemistry, Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal.
i3S, Institute of Research and Innovation in Health, University of Porto, Porto, Portugal.
Cell J. 2021 Jul;23(2):174-183. doi: 10.22074/cellj.2021.7212. Epub 2021 May 26.
The aim is to examine the effect of metformin in human microvascular endothelial cells exposed to high glucose (HG) concentration and compare them with the effects of other 5' adenosine monophosphate-activated protein kinase (AMPK) modulators under the same condition.
In this experimental study, human microvascular endothelial cells (HMECs) were treated with 15 mM metformin, 1 mM 5-aminoimidazol-4-carboxamideribonucleotide (AICAR) and 10 mM compound C in the presence of 20 mM glucose (hyperglycemic condition). Migration, invasion and proliferation were evaluated as well as the capillary-like structures formation. Moreover, the expression of angiogenic genes was assessed.
Metformin significantly inhibited vessel formation and migration, although it did not change HMECs proliferation and invasion. In addition, metformin significantly reduced collagen formation as evidenced by histological staining. Concomitantly, expression of several genes implicated in angiogenesis and fibrosis, namely and , was slightly upregulated. Immunostaining for proteins involved in ALK5 receptor signaling, the alternative TGFß signaling pathway, revealed significant differences in SMAD2/3 expression.
Our data showed that metformin prevents vessel assembly in HMECs, probably through an AMPKindependent mechanism. Understanding the molecular mechanisms by which this pharmacological agent affects endothelial dysfunction is of paramount importance and paves the way to its particular use in preventing development of diabetic retinopathy and nephropathy, two processes where angiogenesis is exacerbated.
研究二甲双胍对暴露于高糖（HG）浓度下的人微血管内皮细胞的影响，并将其与相同条件下其他5'-腺苷单磷酸激活蛋白激酶（AMPK）调节剂的作用进行比较。
在本实验研究中，在20 mM葡萄糖（高血糖条件）存在的情况下，用人微血管内皮细胞（HMECs）分别用15 mM二甲双胍、1 mM 5-氨基咪唑-4-甲酰胺核糖核苷酸（AICAR）和10 mM化合物C进行处理。评估细胞迁移、侵袭和增殖以及毛细血管样结构的形成。此外，还评估了血管生成基因的表达。
二甲双胍显著抑制血管形成和迁移，尽管它没有改变HMECs的增殖和侵袭。此外，组织学染色证明二甲双胍显著减少胶原蛋白的形成。同时，可以看到一些参与血管生成和纤维化的基因，即[具体基因名称未给出]的表达略有上调。对ALK5受体信号通路（替代性TGFβ信号通路）中相关蛋白的免疫染色显示，SMAD2/3表达存在显著差异。
我们的数据表明，二甲双胍可能通过非AMPK依赖机制阻止HMECs中的血管组装。了解这种药物影响内皮功能障碍的分子机制至关重要，并为其在预防糖尿病视网膜病变和肾病（这两个血管生成加剧的过程）发展中的特殊应用铺平了道路。
