Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea.
Department of Internal Medicine, Severance Hospital, Institute of Endocrine Research, Yonsei University College of Medicine, Seoul, Korea.
PLoS One. 2020 Aug 6;15(8):e0237015. doi: 10.1371/journal.pone.0237015. eCollection 2020.
Graves' orbitopathy (GO) is characterised in early stages by orbital fibroblast inflammation, which can be aggravated by oxidative stress and often leads to fibrosis. Protein tyrosine protein 1B (PTP1B) is a regulator of inflammation and a therapeutic target in diabetes. We investigated the role of PTP1B in the GO mechanism using orbital fibroblasts from GO and healthy non-GO subjects. After 24 hours of transfection with PTPN1 siRNA, the fibroblasts were exposed to interleukin (IL)-1β, cigarette smoke extract (CSE), H2O2, and transforming growth factor (TGF)-β stimulations. Inflammatory cytokines and fibrosis-related proteins were analysed using western blotting and/or enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) release was detected using an oxidant-sensitive fluorescent probe. IL-1β, tumor necrosis factor (TNF)-α, bovine thyroid stimulating hormone (bTSH), high-affinity human stimulatory monoclonal antibody of TSH receptor (M22), and insulin-like growth factor-1 (IGF-1) significantly increased PTP1B protein production in GO and non-GO fibroblasts. PTPN1 silencing significantly blocked IL-1β-induced inflammatory cytokine production, CSE- and H2O2-induced ROS synthesis, and TGF-β-induced expression of collagen Iα, α-smooth muscle actin (SMA), and fibronectin in GO fibroblasts. Silencing PTPN1 also decreased phosphorylation levels of Akt, p38, and c-Jun N-terminal kinase (JNK) and endoplasmic reticulum (ER)-stress response proteins in GO cells. PTP1B may be a potential therapeutic target of anti-inflammatory, anti-oxidant and anti-fibrotic treatment of GO.
格雷夫斯眼病（GO）的早期特征是眼眶成纤维细胞炎症，氧化应激可加重这种炎症，进而导致纤维化。蛋白酪氨酸磷酸酶 1B（PTP1B）是炎症的调节剂，也是糖尿病的治疗靶点。我们使用 GO 和非 GO 患者的眼眶成纤维细胞，研究了 PTP1B 在 GO 发病机制中的作用。用 PTPN1 siRNA 转染 24 小时后，将成纤维细胞暴露于白细胞介素（IL）-1β、香烟烟雾提取物（CSE）、H2O2 和转化生长因子（TGF）-β刺激物中。使用 Western blot 和/或酶联免疫吸附测定（ELISA）分析炎症细胞因子和纤维化相关蛋白。使用氧化敏感荧光探针检测活性氧（ROS）释放。IL-1β、肿瘤坏死因子（TNF）-α、牛甲状腺刺激素（bTSH）、高亲和力人 TSH 受体刺激性单克隆抗体（M22）和胰岛素样生长因子-1（IGF-1）显著增加 GO 和非 GO 成纤维细胞中 PTP1B 蛋白的产生。PTPN1 沉默显著阻断了 IL-1β诱导的炎症细胞因子产生、CSE 和 H2O2 诱导的 ROS 合成，以及 TGF-β诱导的 GO 成纤维细胞胶原 Iα、α-平滑肌肌动蛋白（SMA）和纤维连接蛋白的表达。沉默 PTPN1 还降低了 GO 细胞中 Akt、p38 和 c-Jun N-末端激酶（JNK）和内质网（ER）应激反应蛋白的磷酸化水平。PTP1B 可能是 GO 抗炎、抗氧化和抗纤维化治疗的潜在治疗靶点。
