A20/TNFAIP3 Increases ENOS Expression in an ERK5/KLF2-Dependent Manner to Support Endothelial Cell Health in the Face of Inflammation.
A20/TNFAIP3 Increases ENOS Expression in an ERK5/KLF2-Dependent Manner to Support Endothelial Cell Health in the Face of Inflammation.
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A20/TNFAIP3 以 ERK5/KLF2 依赖性方式增加 ENOS 表达，以支持炎症时的内皮细胞健康。
DOI：
10.3389/fcvm.2021.651230
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发表时间：
2021
影响因子：
3.6
通讯作者：
Ferran C
中科院分区：
文献类型：
--
作者：
Angolano C;Kaczmarek E;Essayagh S;Daniel S;Choi LY;Tung B;Sauvage G;Lee A;Kipper FC;Arvelo MB;Moll HP;Ferran C
理论基础：内皮型一氧化氮合酶(ENOS)在炎症和代谢损伤时的表达和活性降低是内皮细胞(EC)功能障碍的标志，它是动脉粥样硬化和高血压发生的前兆。我们之前报道了泛素编辑和抗炎蛋白A20的动脉粥样硬化保护特性，也被称为TNFAIP3，部分是通过阻断EC中的核因子-kappa B(NF-κB)和干扰素信号并保护这些细胞免受凋亡。然而，A20‘S对eNOS表达和功能的影响仍不清楚。在这项研究中，我们评估了A20过度表达或敲除对EC中eNOS表达的影响，在基线和肿瘤坏死因子(TNF)治疗后，用于模拟炎症。方法和结果：人冠状动脉内皮细胞(HCAEC)过表达A20可显著提高基础enos mRNA(QPCR)和蛋白(Western印迹)水平，并可阻止肿瘤坏死因子(TNF)下调其表达。相反，siRNA诱导的A20基因敲除降低了enos的mRNA水平，证明A20是eNOS表达的生理调节因子。通过对人eNOS启动子的缺失和点突变，以及eNOS转录调控因子的敲除，我们证明了20介导的eNOS的增加是转录的，依赖于转录因子Krüppel样因子(KLF2)的表达增加，以及KLF2的上游，依赖于细胞外信号调节激酶5(ERK5)的激活。相应地，ERK5基因被敲除或抑制后，A20‘S上调KLF2和eNOS表达的能力显著减弱。此外，A20在HCAEC中的过表达增加了eNOS Ser-1177的磷酸化，这是该酶功能的关键。结论：A20在EC中的过表达以ERK5/KLF2依赖的方式促进eNOS转录，并促进eNOS激活磷酸化，这是首次报道。这种作用可以抵抗eNOS被肿瘤坏死因子下调的作用，从而在面对炎症时防止EC功能障碍。A20的这一新功能进一步证明了它在预防/治疗动脉粥样硬化方面的治疗前景。
Rationale: Decreased expression and activity of endothelial nitric oxide synthase (eNOS) in response to inflammatory and metabolic insults is the hallmark of endothelial cell (EC) dysfunction that preludes the development of atherosclerosis and hypertension. We previously reported the atheroprotective properties of the ubiquitin-editing and anti-inflammatory protein A20, also known as TNFAIP3, in part through interrupting nuclear factor-kappa B (NF-κB) and interferon signaling in EC and protecting these cells from apoptosis. However, A20's effect on eNOS expression and function remains unknown. In this study, we evaluated the impact of A20 overexpression or knockdown on eNOS expression in EC, at baseline and after tumor necrosis factor (TNF) treatment, used to mimic inflammation. Methods and Results: A20 overexpression in human coronary artery EC (HCAEC) significantly increased basal eNOS mRNA (qPCR) and protein (western blot) levels and prevented their downregulation by TNF. Conversely, siRNA-induced A20 knockdown decreased eNOS mRNA levels, identifying A20 as a physiologic regulator of eNOS expression. By reporter assays, using deletion and point mutants of the human eNOS promoter, and knockdown of eNOS transcriptional regulators, we demonstrated that A20-mediated increase of eNOS was transcriptional and relied on increased expression of the transcription factor Krüppel-like factor (KLF2), and upstream of KLF2, on activation of extracellular signal-regulated kinase 5 (ERK5). Accordingly, ERK5 knockdown or inhibition significantly abrogated A20's ability to increase KLF2 and eNOS expression. In addition, A20 overexpression in HCAEC increased eNOS phosphorylation at Ser-1177, which is key for the function of this enzyme. Conclusions: This is the first report demonstrating that overexpression of A20 in EC increases eNOS transcription in an ERK5/KLF2-dependent manner and promotes eNOS activating phosphorylation. This effect withstands eNOS downregulation by TNF, preventing EC dysfunction in the face of inflammation. This novel function of A20 further qualifies its therapeutic promise to prevent/treat atherosclerosis.
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影响因子：
37.8
作者：
da Silva, Cleide Goncalves;Specht, Anke;Kaczmarek, Elzbieta
通讯作者：
Kaczmarek, Elzbieta
影响因子：
7.7
作者：
Boonyasrisawat, Watip;Eberle, Delphine;Doria, Alessandro
通讯作者：
Doria, Alessandro
影响因子：
20.3
作者：
Dekker, RJ;van Soest, S;Horrevoets, AJG
通讯作者：
Horrevoets, AJG
DOI：
10.1111/j.1549-8719.2010.00071.x
发表时间：
2011-02
期刊：
Microcirculation (New York, N.Y. : 1994)
影响因子：
--
作者：
Clark PR;Jensen TJ;Kluger MS;Morelock M;Hanidu A;Qi Z;Tatake RJ;Pober JS
通讯作者：
Pober JS
增强的NF-κB活性通过2型糖尿病中的PARP-1-，SP-1-和COX-2依赖性机制损害了血管功能。
影响因子：
7.7
作者：
Kassan M;Choi SK;Galán M;Bishop A;Umezawa K;Trebak M;Belmadani S;Matrougui K
通讯作者：
Matrougui K