AMP-Activated Protein Kinase Regulates Endothelial Cell Angiotensin-Converting Enzyme Expression via p53 and the Post-Transcriptional Regulation of microRNA-143/145
AMP-Activated Protein Kinase Regulates Endothelial Cell Angiotensin-Converting Enzyme Expression via p53 and the Post-Transcriptional Regulation of microRNA-143/145
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DOI：
10.1161/circresaha.113.301282
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发表时间：
2013-04-12
影响因子：
20.1
通讯作者：
Fleming, Ingrid
中科院分区：
文献类型：
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作者：
Kohlstedt, Karin;Trouvain, Caroline;Fleming, Ingrid
原理：高血管紧张素转换酶 (ACE) 水平与心血管疾病相关，但对其表达的调节知之甚少。目的：评估调节内皮 ACE 表达的分子机制，重点关注 AMP 激活蛋白激酶 (AMPK) 和 miR-143/145 的作用。方法和结果：剪切应力降低培养的内皮细胞中 ACE 的表达，下调 AMPK α 2 可防止这种效应，但不是 AMPK α 1。AMPK α 2(-/-) 小鼠比野生型同窝小鼠表达更高的 ACE 水平，导致 ACE 底物缓激肽的后肢血管舒张受损。后一种反应在内皮细胞中缺乏 AMPK α2 亚基的动物中也很明显。在培养的内皮细胞中，剪切应力以 AMPK α 2 依赖性方式增加 miR-143/145 水平，并且 miR-143/145 过表达降低 ACE 表达。剪切应力的影响与 miR-143/145 启动子活性和转录的增加无关，但可能归因于 AMPK α 2 对前体 miR-143/145 的转录后调节。AMPK 底物 p53 可以增强多种 microRNA（包括 miR-143/145）的转录后加工。我们发现剪切应力引发 p53（Ser15）的 AMPK α 2 依赖性磷酸化，并且 p53 下调可防止剪切应力诱导的 ACE 表达下降。将链脲佐菌素诱导的小鼠糖尿病作为 AMPK 活性改变的病理生理学模型进行研究。糖尿病增加 AMPK 底物、p53 和乙酰辅酶 A 羧化酶的组织磷酸化，这些变化与 miR-143/145 水平增加和 ACE 表达减少相关。 结论：AMPK α 2 通过 p53 磷酸化和 miR-143/145 上调抑制内皮 ACE 表达。 miR-143/145 的转录后调节可能导致与糖尿病相关的血管并发症。 （Circ Res. 2013；112：1150-1158。）
Rationale: High-angiotensin-converting enzyme (ACE)-levels are associated with cardiovascular disease, but little is known about the regulation of its expression.Objective: To assess the molecular mechanisms regulating endothelial ACE expression focusing on the role of the AMP-activated protein kinase (AMPK) and miR-143/145.Methods and Results: Shear stress decreased ACE expression in cultured endothelial cells, an effect prevented by downregulating AMPK alpha 2 but not AMPK alpha 1. AMPK alpha 2(-/-) mice expressed higher ACE levels than wild-type littermates resulting in impaired hindlimb vasodilatation to the ACE substrate, bradykinin. The latter response was also evident in animals lacking the AMPK alpha 2 subunit only in endothelial cells. In cultured endothelial cells, miR-143/145 levels were increased by shear stress in an AMPK alpha 2-dependent manner, and miR-143/145 overexpression decreased ACE expression. The effect of shear stress was unrelated to an increase in miR-143/145 promoter activity and transcription but could be attributed to post-transcriptional regulation of precursor-miR-143/145 by AMPK alpha 2. The AMPK substrate, p53, can enhance the post-transcriptional processing of several microRNAs, including miR-143/145. We found that shear stress elicited the AMPK alpha 2-dependent phosphorylation of p53 (on Ser15), and that p53 downregulation prevented the shear stress-induced decrease in ACE expression. Streptozotocin-induced diabetes mellitus in mice was studied as a pathophysiological model of altered AMPK activity. Diabetes mellitus increased tissue phosphorylation of the AMPK substrates, p53 and acetyl-coenzyme A carboxylase, changes that correlated with increased miR-143/145 levels and decreased ACE expression.Conclusions: AMPK alpha 2 suppresses endothelial ACE expression via the phosphorylation of p53 and upregulation of miR-143/145. Post-transcriptional regulation of miR-143/145 may contribute to the vascular complications associated with diabetes mellitus. (Circ Res. 2013; 112: 1150-1158.)