PDE4B abrogation extenuates angiotensin II-induced endothelial dysfunction related to hypertension through up-regulation of AMPK/Sirt1/Nrf2/ARE signaling

IF 2.7 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2024-11-23 DOI:10.1016/j.tice.2024.102637

Yong Chen, Suipeng Li, Xuqing Hou, Yinfeng Jia

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引用次数: 0

Abstract

Endothelial dysfunction is commonly perceived as a precursor in the process of hypertension, a severe cardiovascular disorder. Phosphodiesterase 4B (PDE4B) inactivation has been proposed to exert cardioprotective effects and prevent pulmonary hypertension. However, the role of PDE4B in endothelial dysfunction in hypertension remains inexplicit, which will be investigated in the present work. In angiotensin II (Ang II)-induced human umbilical vein endothelial cells (HUVECs), RT-qPCR and Western blotting were used to analyze PDE4B expression. CCK-8 method was used to detect cell viability. Flow cytometry assay and Caspase 3 assay kit were used to detect cellular apoptotic level. Wound healing and tube formation assays were respectively used to detect cell migration and angiogenesis. Western blotting and corresponding assay kits were respectively used to analyze the expressions and contents of endothelial dysfunction markers. JC-1 assay, RT-qPCR and relevant assay kit were respectively used to detect mitochondrial membrane potential (ΔΨm), quantify mitochondrial DNA (mtDNA) copy number and mitochondrial permeability transition pore (mPTP) opening. Besides, Western blotting was used to analyze the expressions of endoplasmic reticulum stress (ERS) and AMP-activated protein kinase (AMPK)/sirtuin 1 (Sirt1)/nuclear factor-erythroid 2 related factor 2 (Nrf2)/antioxidant response element (ARE) signaling-associated proteins. PDE4B expression was increased in Ang II- induced HUVECs. PDE4B knockdown promoted the viability, migration, angiogenesis while inhibiting the apoptosis, endothelial dysfunction, ERS and mitochondrial damage in Ang II-induced HUVECs. Additionally, PDE4B silence activated AMPK/Sirt1/Nrf2/ARE pathway and AMPK inhibitor Compound C (CC) partially reversed the effects of PDE4B down-regulation on Ang II-induced HUVECs. Conclusively, PDE4B inhibition might protect against Ang II-induced endothelial dysfunction in HUVECs via up-regulating AMPK/Sirt1/Nrf2/ARE pathway, which might be mediated by the suppression of ERS and mitochondrial damage.

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通过上调AMPK/Sirt1/Nrf2/ARE信号，消减PDE4B可减轻血管紧张素II诱导的与高血压有关的内皮功能障碍

内皮功能障碍通常被认为是高血压这一严重心血管疾病的前兆。磷酸二酯酶 4B (PDE4B) 失活被认为具有保护心脏和预防肺动脉高压的作用。然而，PDE4B 在高血压内皮功能障碍中的作用尚不明确，本研究将对此进行研究。在血管紧张素 II（Ang II）诱导的人脐静脉内皮细胞（HUVECs）中，采用 RT-qPCR 和 Western 印迹法分析 PDE4B 的表达。采用 CCK-8 法检测细胞活力。流式细胞术检测和 Caspase 3 检测试剂盒用于检测细胞凋亡水平。伤口愈合和管形成试验分别用于检测细胞迁移和血管生成。Western 印迹法和相应的检测试剂盒分别用于分析内皮功能障碍标志物的表达和含量。JC-1检测、RT-qPCR和相关检测试剂盒分别用于检测线粒体膜电位（ΔΨm）、线粒体DNA（mtDNA）拷贝数定量和线粒体通透性转换孔（mPTP）开放。此外，还采用 Western 印迹法分析了内质网应激（ERS）和 AMPK 激活蛋白激酶（AMPK）/sirtuin 1（Sirt1）/核因子红细胞 2 相关因子 2（Nrf2）/抗氧化反应元件（ARE）信号相关蛋白的表达。在 Ang II 诱导的 HUVECs 中，PDE4B 的表达增加。PDE4B 敲除可促进 Ang II 诱导的 HUVECs 的活力、迁移和血管生成，同时抑制其凋亡、内皮功能障碍、ERS 和线粒体损伤。此外，PDE4B 沉默激活了 AMPK/Sirt1/Nrf2/ARE 通路，AMPK 抑制剂化合物 C（CC）部分逆转了 PDE4B 下调对 Ang II 诱导的 HUVECs 的影响。结论是，PDE4B抑制可通过上调AMPK/Sirt1/Nrf2/ARE通路保护HUVECs免受Ang II诱导的内皮功能障碍，而AMPK/Sirt1/Nrf2/ARE通路可能是通过抑制ERS和线粒体损伤介导的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.