Proangiogenic effect of thyrotropin receptor stimulating antibody in human umbilical vein endothelial cells.

IF 3 3区医学 Q2 ENDOCRINOLOGY & METABOLISM

Endocrine Pub Date : 2024-10-25 DOI:10.1007/s12020-024-04048-z

Yue Yuan, Xingjia Li, Wenjing Ni, Wenbin Huang, Guofang Chen, Shuhang Xu, Chao Liu

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引用次数: 0

Abstract

Objective: This study aims to investigate the role of TRAb in the angiogenesis associated with Graves' disease (GD) and to elucidate its underlying mechanisms.

Methods: Human thyroid follicular epithelial cells (Nthy-ori 3-1) and human umbilical vein endothelial cells (HUVECs) were treated with the monoclonal thyroid-stimulating antibody M22 and thyroid-stimulating hormone (TSH) at various concentrations. Cell viability, migration, and tube formation were evaluated using CCK-8, wound healing, and tube formation assays, respectively. Protein expressions of TSHR receptor (TSHR) and phosphorylated AKT (p-AKT) in M22-induced HUVECs were quantified via Western blotting. Proteomic analysis was employed to identify changes in protein expression profiles and relevant signaling pathways in GD specimens. Immunofluorescence assays were conducted to detect and localize the expressions of CD34 and PROX1 in GD specimens and normal thyroid tissues.

Results: M22 stimulated the proliferation of Nthy-ori 3-1 cells and HUVECs in a dose-dependent manner, while TSH exhibited an inverted U-shaped dose-response effect. M22 also dose-dependently promoted angiogenesis, and more effectively induced tube formation in HUVECs compared to TSH, although the difference was not statistically significant. A total of 16 proteins were significantly upregulated and 24 were downregulated in M22-induced HUVECs. Notably, PROX1, the most significantly upregulated protein, is closely associated with angiogenesis. Immunofluorescence confirmed that PROX1 was significantly more expressed in thyroid tissues from GD patients compared to normal tissues adjacent to papillary thyroid cancer (PTC), and it co-localized with CD34.

Conclusion: TRAb enhances angiogenesis and upregulates PROX1 expression in HUVECs, suggesting a novel possible mechanism for goiter formation in GD.

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促甲状腺激素受体刺激抗体在人脐静脉内皮细胞中的促血管生成作用。

研究目的方法：用不同浓度的单克隆促甲状腺抗体M22和促甲状腺激素（TSH）处理人甲状腺滤泡上皮细胞（Nthy-ori 3-1）和人脐静脉内皮细胞（HUVECs）。分别使用 CCK-8、伤口愈合和管形成试验评估细胞活力、迁移和管形成。通过 Western 印迹对 M22 诱导的 HUVEC 中 TSHR 受体（TSHR）和磷酸化 AKT（p-AKT）的蛋白表达进行量化。蛋白质组分析用于确定广东样本中蛋白质表达谱的变化和相关信号通路。免疫荧光试验检测了CD34和PROX1在GD标本和正常甲状腺组织中的表达：结果：M22以剂量依赖的方式刺激Nthy-ori 3-1细胞和HUVEC的增殖，而TSH表现出倒U形的剂量反应效应。M22 对血管生成也有剂量依赖性的促进作用，与 TSH 相比，M22 能更有效地诱导 HUVECs 中血管管的形成，但差异无统计学意义。在 M22 诱导的 HUVECs 中，共有 16 种蛋白质被显著上调，24 种被下调。值得注意的是，上调最明显的蛋白 PROX1 与血管生成密切相关。免疫荧光证实，与邻近甲状腺乳头状癌（PTC）的正常组织相比，PROX1在GD患者甲状腺组织中的表达明显增加，并且与CD34共定位：结论：TRAb可促进血管生成并上调HUVECs中PROX1的表达，这可能是GD患者甲状腺肿形成的一种新机制。

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

Endocrine ENDOCRINOLOGY & METABOLISM-

CiteScore

6.50

自引率

5.40%

发文量

295

审稿时长

1.5 months

期刊介绍： Well-established as a major journal in today’s rapidly advancing experimental and clinical research areas, Endocrine publishes original articles devoted to basic (including molecular, cellular and physiological studies), translational and clinical research in all the different fields of endocrinology and metabolism. Articles will be accepted based on peer-reviews, priority, and editorial decision. Invited reviews, mini-reviews and viewpoints on relevant pathophysiological and clinical topics, as well as Editorials on articles appearing in the Journal, are published. Unsolicited Editorials will be evaluated by the editorial team. Outcomes of scientific meetings, as well as guidelines and position statements, may be submitted. The Journal also considers special feature articles in the field of endocrine genetics and epigenetics, as well as articles devoted to novel methods and techniques in endocrinology. Endocrine covers controversial, clinical endocrine issues. Meta-analyses on endocrine and metabolic topics are also accepted. Descriptions of single clinical cases and/or small patients studies are not published unless of exceptional interest. However, reports of novel imaging studies and endocrine side effects in single patients may be considered. Research letters and letters to the editor related or unrelated to recently published articles can be submitted. Endocrine covers leading topics in endocrinology such as neuroendocrinology, pituitary and hypothalamic peptides, thyroid physiological and clinical aspects, bone and mineral metabolism and osteoporosis, obesity, lipid and energy metabolism and food intake control, insulin, Type 1 and Type 2 diabetes, hormones of male and female reproduction, adrenal diseases pediatric and geriatric endocrinology, endocrine hypertension and endocrine oncology.