Division Department of Cardiology Organization, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Key Laboratories of the Education Ministry for Myocardial Ischemia Mechanisms and Treatment, Harbin, China.
J Transl Med. 2019 Nov 15;17(1):378. doi: 10.1186/s12967-019-02135-8.
Atherosclerosis preferentially develops in regions of disturbed flow (DF). Emerging evidence indicates that yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ), which are both effectors of the Hippo pathway, sense different blood flow patterns and regulate atherosclerotic lesions. We previously found that methotrexate (MTX) reduces in-stent neoatherosclerosis, decreases the plaque burden, and has an effect on local fluid shear stress. Here, we investigated the atheroprotective effect of MTX under DF and the mechanisms underlying these properties.
Human umbilical vein endothelial cells (HUVECs) were subjected to biomechanical stretch using a parallel-plate flow system and treated with or without MTX at therapeutically relevant concentrations. Additionally, an extravascular device was used to induce DF in the left common carotid artery of C57BL/6 mice, followed by treatment with MTX or 0.9% saline. The artery was then assessed histopathologically after 4 weeks on a Western diet.
We observed that MTX significantly inhibited DF-induced endothelial YAP/TAZ activation. Furthermore, it markedly decreased pro-inflammatory factor secretion and monocyte adhesion in HUVECs but had no effect on apoptosis. Mechanistically, AMPKa1 depletion attenuated these effects of MTX. Accordingly, MTX decreased DF-induced plaque formation, which was accompanied by YAP/TAZ downregulation in vivo.
Taken together, we conclude that MTX exerts protective effects via the AMP-dependent kinase (AMPK)-YAP/TAZ pathway. These results provide a basis for the prevention and treatment of atherosclerosis via the inhibition of YAP/TAZ.
动脉粥样硬化优先在血流紊乱（DF）的区域发展。新出现的证据表明，Yes 相关蛋白（YAP）和转录共激活因子与 PDZ 结合基序（TAZ），它们都是 Hippo 通路的效应物，感知不同的血流模式并调节动脉粥样硬化病变。我们之前发现甲氨蝶呤（MTX）可减少支架内新生动脉粥样硬化，减少斑块负担，并对局部流体剪切力产生影响。在这里，我们研究了 MTX 在 DF 下的抗动脉粥样硬化作用及其潜在机制。
采用平行平板流动系统对人脐静脉内皮细胞（HUVEC）进行生物力学拉伸，并在治疗相关浓度下用或不用 MTX 处理。此外，还使用血管外装置在 C57BL/6 小鼠的左颈总动脉上诱导 DF，然后用 MTX 或 0.9%生理盐水处理。在西方饮食 4 周后，对动脉进行组织病理学评估。
我们观察到 MTX 显著抑制了 DF 诱导的内皮 YAP/TAZ 激活。此外，它显著降低了 HUVEC 中促炎因子的分泌和单核细胞的黏附，但对细胞凋亡没有影响。机制上，AMPKa1 耗竭减弱了 MTX 的这些作用。因此，MTX 减少了 DF 诱导的斑块形成，体内 YAP/TAZ 下调。
综上所述，我们得出结论，MTX 通过 AMP 依赖性激酶（AMPK）-YAP/TAZ 通路发挥保护作用。这些结果为通过抑制 YAP/TAZ 预防和治疗动脉粥样硬化提供了依据。
