Pole of Pharmacology and Therapeutics, Institute of Experimental and Clinical Research (IREC), Université Catholique de Louvain (UCLouvain), B-1200 Brussels, Belgium.
Pole of Morphology, Institute of Experimental and Clinical Research (IREC), UCLouvain, B-1200 Brussels, Belgium.
Int J Mol Sci. 2021 Dec 23;23(1):153. doi: 10.3390/ijms23010153.
Graves' disease (GD) is an autoimmune thyroiditis often associated with Graves' orbitopathy (GO). GD thyroid and GO orbital fat share high oxidative stress (OS) and hypervascularization. We investigated the metabolic pathways leading to OS and angiogenesis, aiming to further decipher the link between local and systemic GD manifestations. Plasma and thyroid samples were obtained from patients operated on for multinodular goiters (controls) or GD. Orbital fats were from GO or control patients. The NADPH-oxidase-4 (NOX4)/HIF-1α/VEGF-A signaling pathway was investigated by Western blotting and immunostaining. miR-199a family expression was evaluated following quantitative real-time PCR and/or in situ hybridization. In GD thyroids and GO orbital fats, NOX4 was upregulated and correlated with HIF-1α stabilization and VEGF-A overexpression. The biotin assay identified NOX4, HIF-1α and VEGF-A as direct targets of miR-199a-5p in cultured thyrocytes. Interestingly, GD thyroids, GD plasmas and GO orbital fats showed a downregulation of miR-199a-3p/-5p. Our results also highlighted an activation of STAT-3 signaling in GD thyroids and GO orbital fats, a transcription factor known to negatively regulate miR-199a expression. We identified NOX4/HIF-1α/VEGF-A as critical actors in GD and GO. STAT-3-dependent regulation of miR-199a is proposed as a common driver leading to these events in GD thyroids and GO orbital fats.
格雷夫斯病（GD）是一种自身免疫性甲状腺炎，常伴有格雷夫斯眼病（GO）。GD 甲状腺和 GO 眼眶脂肪均存在高氧化应激（OS）和高血管生成。我们研究了导致 OS 和血管生成的代谢途径，旨在进一步阐明 GD 局部和全身表现之间的联系。从接受多结节性甲状腺肿（对照组）或 GD 手术的患者中获得血浆和甲状腺样本。眼眶脂肪来自 GO 或对照组患者。通过 Western blot 和免疫染色研究 NADPH 氧化酶-4（NOX4）/HIF-1α/VEGF-A 信号通路。通过定量实时 PCR 和/或原位杂交评估 miR-199a 家族的表达。在 GD 甲状腺和 GO 眼眶脂肪中，NOX4 上调，并与 HIF-1α稳定和 VEGF-A 过表达相关。生物素测定鉴定出 NOX4、HIF-1α 和 VEGF-A 是培养甲状腺细胞中 miR-199a-5p 的直接靶标。有趣的是，GD 甲状腺、GD 血浆和 GO 眼眶脂肪中 miR-199a-3p/-5p 下调。我们的研究结果还强调了 GD 甲状腺和 GO 眼眶脂肪中 STAT-3 信号的激活，该转录因子已知可负调控 miR-199a 的表达。我们确定了 NOX4/HIF-1α/VEGF-A 是 GD 和 GO 的关键因素。提出 STAT-3 依赖性调节 miR-199a 是导致 GD 甲状腺和 GO 眼眶脂肪中这些事件的共同驱动因素。
