Deregulation of microRNA-503 Contributes to Diabetes Mellitus-Induced Impairment of Endothelial Function and Reparative Angiogenesis After Limb Ischemia
Deregulation of microRNA-503 Contributes to Diabetes Mellitus-Induced Impairment of Endothelial Function and Reparative Angiogenesis After Limb Ischemia
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DOI：
10.1161/circulationaha.110.952325
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发表时间：
2011-01-25
期刊：
影响因子：
37.8
通讯作者：
Emanueli, Costanza
中科院分区：
文献类型：
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作者：
Caporali, Andrea;Meloni, Marco;Emanueli, Costanza
背景-糖尿病损害内皮细胞（EC）功能和缺血后修复性新血管形成的分子机制尚不完全清楚。microRNA负调控靶基因的表达主要是通过在其3'非翻译区的相互作用。方法和结果-我们发现在模拟糖尿病（高D-葡萄糖）和缺血相关饥饿（低生长因子）的培养条件下，内皮细胞中的microRNA-503（miR-503）表达上调。在正常培养条件下，慢病毒介导的miR-503强制表达抑制EC增殖、迁移和在Matrigel上的网络形成（与慢病毒比较）。GFP对照）。相反，通过腺病毒介导的miR-503诱饵转移（Ad.decoymiR-503）或通过antimiR-503（反义寡核苷酸）阻断miR-503活性可改善在高D-葡萄糖/低生长因子条件下培养的EC的功能能力。我们鉴定了CCNE 1和cdc 25 A作为miR-503的直接靶点，其在EC中被高糖/低生长因子下调。接下来，我们获得的证据表明，miR-503的表达在链脲佐菌素糖尿病小鼠的缺血肢体肌肉和从这些肌肉中富集的EC中增加。此外，向糖尿病小鼠的缺血内收肌递送Ad.decoymiR-503纠正了糖尿病诱导的缺血后血管生成和血流恢复的损害。最后，我们研究了miR-503和靶基因在糖尿病患者截肢缺血腿肌肉标本中的表达。作为对照，非糖尿病和非缺血性患者接受隐静脉剥脱的小腿活检。在糖尿病肌肉中，miR-503的表达显著升高，并且与cdc 25蛋白表达呈负相关。血浆miR-503水平也升高，在糖尿病individual. Conclusions，我们的数据表明，miR-503作为一个可能的治疗目标，在糖尿病患者的严重肢体缺血。（循环。2011; 123：282-291）。
Background-Diabetes mellitus impairs endothelial cell (EC) function and postischemic reparative neovascularization by molecular mechanisms that are not fully understood. microRNAs negatively regulate the expression of target genes mainly by interaction in their 3' untranslated region.Methods and Results-We found that microRNA-503 (miR-503) expression in ECs is upregulated in culture conditions mimicking diabetes mellitus (high D-glucose) and ischemia-associated starvation (low growth factors). Under normal culture conditions, lentivirus-mediated miR-503-forced expression inhibited EC proliferation, migration, and network formation on Matrigel (comparisons versus lentivirus. GFP control). Conversely, blocking miR-503 activity by either adenovirus-mediated transfer of a miR-503 decoy (Ad.decoymiR-503) or by antimiR-503 (antisense oligonucleotide) improved the functional capacities of ECs cultured under high D-glucose/low growth factors. We identified CCNE1 and cdc25A as direct miR-503 targets which are downregulated by high glucose/low growth factors in ECs. Next, we obtained evidence that miR-503 expression is increased in ischemic limb muscles of streptozotocin-diabetic mice and in ECs enriched from these muscles. Moreover, Ad.decoymiR-503 delivery to the ischemic adductor of diabetic mice corrected diabetes mellitus-induced impairment of postischemic angiogenesis and blood flow recovery. We finally investigated miR-503 and target gene expression in muscular specimens from the amputated ischemic legs of diabetic patients. As controls, calf biopsies of nondiabetic and nonischemic patients undergoing saphenous vein stripping were used. In diabetic muscles, miR-503 expression was remarkably higher, and it inversely correlated with cdc25 protein expression. Plasma miR-503 levels were also elevated in the diabetic individuals.Conclusions-Our data suggest miR-503 as a possible therapeutic target in diabetic patients with critical limb ischemia. (Circulation. 2011; 123: 282-291.)