Department of Ophthalmology, Soonchunhyang University Bucheon Hospital, Soonchunhyang University College of Medicine, Bucheon, Republic of Korea.
Department of Ophthalmology, Severance Hospital, The Institute of Vision Research, Yonsei University College of Medicine, Seoul, Republic of Korea.
PLoS One. 2022 Aug 18;17(8):e0270416. doi: 10.1371/journal.pone.0270416. eCollection 2022.
To investigate the role of microRNA (miR)-155 in inflammation in an in-vitro model of Graves' orbitopathy (GO). The expression levels of miR-155 were compared between GO and non-GO orbital tissues. The effects of inflammatory stimulation of interleukin (IL)-1β and tumour necrosis factor alpha (TNF-α) on miR-155 expression on GO and non-GO orbital fibroblasts (OFs) were investigated. The effects of miR-155 mimics and inhibitors of inflammatory proteins and IL-2-inducible T-cell kinase (ITK) expression were examined, along with those related to the knockdown of ITK with siITK transfection on inflammatory proteins. We also examined how ITK inhibitors affect miR-155 expression in GO and non-GO OFs. The expression levels of miR-155 were higher in GO orbital tissues than in non-GO tissue. The overexpression of miR-155 was induced by IL-1β and TNF-α in OFs from GO and non-GO patients. IL-1β-induced IL-6 (ICAM1) protein production was significantly reduced (increased) by miR-155 mimics and inhibitors. The mRNA and protein levels of ITK were downregulated by overexpressed miR-155 via miR-155 mimics. Knockdown of ITK via siITK transfection induced a decrease in the expression levels of ITK, IL-17, IL-6, IL-1β, and TNF-α protein. The expression of miR-155 was significantly downregulated by treatment with ITK inhibitors and Bruton's tyrosine kinase (BTK)/ITK dual inhibitors in a time-dependent manner. Our results indicated a potential relationship between miR-155 and ITK in the context of GO OFs. The overexpression of miR-155 repressed ITK expression and relieved inflammation. Thus, miR-155 appears to have anti-inflammatory effects in GO OFs. This discovery provides a new concept for developing GO treatment therapeutics.
为了探讨微小 RNA（miR）-155 在甲状腺眼病（GO）体外模型中的炎症作用。比较 GO 和非 GO 眼眶组织中 miR-155 的表达水平。研究了白细胞介素（IL）-1β和肿瘤坏死因子-α（TNF-α）对 GO 和非 GO 眼眶成纤维细胞（OFs）中 miR-155 表达的炎症刺激作用。检测了炎症蛋白和 IL-2 诱导的 T 细胞激酶（ITK）表达的 miR-155 模拟物和抑制剂的作用，以及用 siITK 转染对炎症蛋白进行 ITK 敲低的作用。我们还研究了 ITK 抑制剂如何影响 GO 和非 GO OFs 中的 miR-155 表达。GO 眼眶组织中的 miR-155 表达水平高于非 GO 组织。IL-1β 和 TNF-α 在 GO 和非 GO 患者的 OFs 中诱导 miR-155 的过表达。miR-155 模拟物和抑制剂显著降低（增加）了 IL-1β 诱导的 IL-6（ICAM1）蛋白的产生。通过 miR-155 模拟物过表达 miR-155 下调 ITK 的 mRNA 和蛋白水平。通过 siITK 转染敲低 ITK 诱导 ITK、IL-17、IL-6、IL-1β 和 TNF-α 蛋白的表达水平降低。用 ITK 抑制剂和 Bruton 酪氨酸激酶（BTK）/ITK 双重抑制剂处理可显著下调 miR-155 的表达，呈时间依赖性。我们的结果表明，miR-155 与 GO OFs 中的 ITK 之间存在潜在关系。miR-155 的过表达抑制 ITK 的表达并缓解炎症。因此，miR-155 在 GO OFs 中具有抗炎作用。这一发现为 GO 治疗提供了一个新的概念。
