High glucose increases the formation and pro-oxidative activity of endothelial microparticles
High glucose increases the formation and pro-oxidative activity of endothelial microparticles
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DOI：
10.1007/s00125-017-4331-2
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发表时间：
2017-09-01
期刊：
影响因子：
8.2
通讯作者：
Burns, Kevin D.
中科院分区：
文献类型：
--
作者：
Burger, Dylan;Turner, Maddison;Burns, Kevin D.
糖尿病患者表现出循环内皮微粒（eMP，也称为内皮微泡）的增加，这与内皮功能障碍和心血管并发症的风险增加有关。我们已经表明，虽然他们在糖尿病中的作用还不清楚，但他们是血管损伤的标志物和介质。我们假设，组成和EMPs的生物活性改变，在响应高葡萄糖暴露。方法从正常葡萄糖（NG）、高糖（HG）和L-葡萄糖渗透压控制（LG）条件下培养的人脐静脉内皮细胞（HUVECs）培养液中分离eMP。通过纳米颗粒追踪分析和流式细胞术评估eMP的尺寸、浓度和表面电荷。结果高糖培养24 h后，内皮细胞内eMP形成量增加3倍，平均粒径增大（269 +/- 18 nm vs 226 +/- 11 nm），表面电荷减少（P < 0. 05）。与eMP（NG）或eMP（LG）相比，eMP（HG）具有约3倍的促凝血活性，更大程度地刺激HUVEC ROS产生（与eMP（NG）的250%相似），并且是更有效的内皮依赖性舒张抑制剂。eMP的蛋白质组学分析确定了1212个独立的蛋白质，其中68个是专门发现在eMP（HG）。基因本体论分析表明，eMP（HG）-排他性的蛋白质与相关的血液凝固，细胞信号和免疫细胞activation.Conclusions/interpretation的信号通路我们的研究结果表明，葡萄糖升高是一个有力的刺激eMP形成，也改变了它们的分子组成，导致生物活性增加。这些影响可能导致糖尿病患者进行性内皮损伤和随后的心血管并发症。
Aims/hypothesis Individuals with diabetes exhibit increases in circulating endothelial microparticles (eMPs, also referred to as endothelial microvesicles), which are associated with endothelial dysfunction and a heightened risk of cardiovascular complications. We have shown that eMPs are markers and mediators of vascular injury although their role in diabetes is unclear. We hypothesised that the composition and biological activity of eMPs are altered in response to high glucose exposure. We assessed the effects of high glucose on eMP formation, composition and signalling in cultured HUVECs.Methods eMPs were isolated from the media of HUVECs cultured under conditions of normal glucose (eMP(NG)), high glucose (eMP(HG)) or osmotic control of L-glucose (eMP(LG)). eMP size, concentration and surface charge were assessed by nanoparticle tracking analysis and flow cytometry. eMP protein composition was assessed by liquid chromatography-tandem mass spectrometry, and eMP-mediated effects on coagulation, reactive oxygen species (ROS) production and vessel function were assessed.Results Exposure of HUVECs to high glucose for 24 h caused a threefold increase in eMP formation, increased mean particle size (269 +/- 18 nm vs 226 +/- 11 nm) and decreased surface charge. Compared with eMP(NG) or eMP(LG), eMP(HG) possessed approximately threefold greater pro-coagulant activity, stimulated HUVEC ROS production to a greater extent (similar to 250% of eMP(NG)) and were more potent inhibitors of endothelial-dependent relaxation. Proteomic analysis of eMPs identified 1212 independent proteins of which 68 were exclusively found in eMP(HG). Gene ontology analysis revealed that eMP(HG)-exclusive proteins were associated with signalling pathways related to blood coagulation, cell signalling and immune cell activation.Conclusions/interpretation Our results indicate that elevated glucose is a potent stimulus for eMP formation that also alters their molecular composition leading to increased bioactivity. Such effects may contribute to progressive endothelial injury and subsequent cardiovascular complications in diabetes.