AMP-activated Protein Kinase Phosphorylation of Angiotensin-Converting Enzyme 2 in Endothelium Mitigates Pulmonary Hypertension
AMP-activated Protein Kinase Phosphorylation of Angiotensin-Converting Enzyme 2 in Endothelium Mitigates Pulmonary Hypertension
复制标题
内皮细胞中 AMP 激活的蛋白激酶磷酸化血管紧张素转换酶 2 可减轻肺动脉高压
DOI：
10.1164/rccm.201712-2570oc
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发表时间：
2018-08-15
影响因子：
24.7
通讯作者：
Shyy, John Y. -J.
中科院分区：
文献类型：
--
作者：
Zhang, Jiao;Dong, Jianjie;Shyy, John Y. -J.
原理：内皮功能障碍在肺动脉高压（PH）中起着不可或缺的作用。AMPK（AMP活化蛋白激酶）和ACE 2（血管紧张素转换酶2）在内皮稳态中至关重要。AMPK调节肺内皮中ACE 2的机制及其在PH中的保护作用仍然难以捉摸。目的：我们研究了ACE 2 Ser 680的AMPK磷酸化在ACE 2稳定性中的作用，并破译了ACE 2的这种翻译后修饰在内皮细胞中的功能后果稳态和PH。方法：应用生物信息学预测、激酶活性测定和磷酸化ACE 2 Ser 680抗体（p-ACE 2 S680）研究AMPK磷酸化ACE 2 Ser 680在内皮细胞中的作用。使用CRISPR-Cas9基因组编辑，我们创建了功能获得型ACE 2 S680 D敲入和功能丧失型ACE 2敲除（ACE 22/2）小鼠系，以解决p-ACE 2 S680和ACE 2参与PH的问题。AMPK-p-ACE 2 S680轴也在来自患有特发性肺动脉高压的人的肺组织中得到验证。AMPK对ACE 2的磷酸化增强了ACE 2的稳定性，从而增加了Ang（血管紧张素）1-7和内皮型一氧化氮合酶衍生的NO的生物利用度。与野生型同窝小鼠相比，ACE 2 S680 D基因敲入小鼠对PH具有抗性。相反，ACE 2敲除小鼠加重了PH，这是在AMPK α 2内皮细胞特异性缺失的小鼠中发现的类似表型。结论：AMPK对肺血管内皮细胞ACE 2 Ser 680磷酸化的影响导致ACE 2不稳定，从而导致肺动脉高压的发生，AMPK对ACE 2的调节是肺动脉高压治疗的潜在靶点。
Rationale: Endothelial dysfunction plays an integral role in pulmonary hypertension (PH). AMPK (AMP-activated protein kinase) and ACE2 (angiotensin-converting enzyme 2) are crucial in endothelial homeostasis. The mechanism by which AMPK regulates ACE2 in the pulmonary endothelium and its protective role in PH remain elusive.Objectives: We investigated the role of AMPK phosphorylation of ACE2 Ser680 in ACE2 stability and deciphered the functional consequences of this post-translational modification of ACE2 in endothelial homeostasis and PH.Methods: Bioinformatics prediction, kinase assay, and antibody against phospho-ACE2 Ser680 (p-ACE2 S680) were used to investigate AMPK phosphorylation of ACE2 Ser680 in endothelial cells. Using CRISPR-Cas9 genomic editing, we created gain-of-function ACE2 S680D knock-in and loss-of-function ACE2 knockout (ACE22/2) mouse lines to address the involvement of p-ACE2 S680 and ACE2 in PH. The AMPK-p-ACE2 S680 axis was also validated in lung tissue from humans with idiopathic pulmonary arterial hypertension.Measurements and Main Results: Phosphorylation of ACE2 by AMPK enhanced the stability of ACE2, which increased Ang (angiotensin) 1-7 and endothelial nitric oxide synthase-derived NO bioavailability. ACE2 S680D knock-in mice were resistant to PH as compared with wild-type littermates. In contrast, ACE2-knockout mice exacerbated PH, a similar phenotype found in mice with endothelial cell-specific deletion of AMPK alpha 2. Consistently, the concentrations of phosphorylated AMPK, p-ACE2 S680, and ACE2 were decreased in human lungs with idiopathic pulmonary arterial hypertension.Conclusions: Impaired phosphorylation of ACE2 Ser680 by AMPK in pulmonary endothelium leads to a labile ACE2 and hence is associated with the pathogenesis of PH. Thus, AMPK regulation of the vasoprotective ACE2 is a potential target for PH treatment.