Hyperglycemia and Advanced Glycation End Products Regulate miR-126 Expression in Endothelial Progenitor Cells
Hyperglycemia and Advanced Glycation End Products Regulate miR-126 Expression in Endothelial Progenitor Cells
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高血糖和晚期糖基化终产物调节内皮祖细胞中的 miR-126 表达
DOI：
10.1159/000448713
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发表时间：
2016-01-01
影响因子：
1.7
通讯作者：
Meng, Shu
中科院分区：
文献类型：
--
作者：
Li, Yeting;Zhou, Qing;Meng, Shu
背景/目的：内皮祖细胞（EPCs）功能障碍与糖尿病血管疾病有关。我们报道了糖尿病患者miR-126下调导致EPC功能障碍。该研究旨在探讨高糖（HG）和晚期糖基化终产物（AGEs）如何调节miR-126的表达，以及miR-126是否介导HG和AGEs对EPCs的影响。方法：先检测葡萄糖（5.5 ~ 50 mM）和AGEs （50 ~ 200 mg/l）对EPC增殖的影响，选择HG （50 mM）和AGEs （50 mg/l）进一步实验。用HG和AGEs刺激EPCs， real-time PCR检测miR-126的表达。采用免疫荧光显微镜和流式细胞术检测活性氧（ROS）。ELISA法检测EPC上清液中IL-6、TNF-α水平。我们还评估了HG和AGEs刺激下miR-126对ROS和炎症标志物的影响。最后，我们检测了PI3K和Akt抑制剂对年龄介导的miR-126表达的影响。结果：HG和AGEs增加了EPCs中IL-6、TNF-α和ROS的表达，降低了miR-126的表达。miR-126负调控IL-6、TNF-α和ROS。miR-126过表达降低，miR-126抑制进一步增加HG和AGEs诱导的炎症标志物和ROS。在age存在的情况下，PI3K和Akt抑制剂进一步降低miR-126的表达。结论：高血糖和AGEs可降低EPCs中miR-126的表达。恢复miR-126表达可能保护EPCs免受HG和AGEs诱导的功能障碍。
Background/Aims: Dysfunction of endothelial progenitor cell (EPCs) contributes to diabetic vascular disease. We reported that downregulated miR-126 in diabetic patients causes EPC dysfunction. The study was designed to investigate how high glucose (HG) and advanced glycation end products (AGEs) regulate miR-126 expression and whether miR-126 mediates the effects of HG and AGEs on EPCs. Methods: We first tested the effects of glucose (5.5-50 mM) and AGEs at 50-200 mg/l on EPC proliferation and selected HG at 50 mM and AGEs at 50 mg/l for further experiments. EPCs were stimulated with HG and AGEs, and miR-126 expression was measured by real-time PCR. Reactive oxygen species (ROS) were measured by immunofluorescence microscopy and flow cytometry. IL-6 and TNF-α levels in EPC supernatants were determined by ELISA. The effects of miR-126 on ROS and inflammatory markers under stimulation of HG and AGEs were also assessed. Finally, the effects of inhibitors of PI3K and Akt on AGE-mediated miR-126 expression were examined. Results: HG and AGEs increased IL-6, TNF-α and ROS and decreased miR-126 expression in EPCs. miR-126 negatively regulated IL-6, TNF-α and ROS. miR-126 overexpression reduced and miR-126 inhibition further increased the inflammatory markers and ROS induced by HG and AGEs. Inhibitors of PI3K and Akt further decreased miR-126 expression in the presence of AGEs. Conclusions: In conclusion, hyperglycemia and AGEs decrease miR-126 expression in EPCs. Recovering miR-126 expression may protect EPCs against dysfunction induced by HG and AGEs.