Liu Jin, Zhu Tianyi, Yang Li, Qian Weijin, Fang Lianfei, Zhang Weiqi, Zhang Haiyang, Wang Yi, Yu Baiguang, Sun Jing, Li Bin, Li Dan, Li Yinwei, Fang Sijie, Zhou Huifang
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, and Center for Basic Medical Research and Innovation in Visual System Diseases of Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Immunology and Microbiology, Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Thyroid. 2025 Jul;35(7):803-815. doi: 10.1089/thy.2025.0062. Epub 2025 Jun 12.
Thyroid eye disease (TED) is a sight-threatening autoimmune disease with cigarette smoking as one of the key risk factors. Cigarette smoking affects both the severity of TED and the patient's response to medication. However, the underlying pathogenic mechanisms of smoking in TED remain unclear. Orbital fibroblasts (OFs) were extracted from patients with TED and non-TED controls, and treated with cigarette smoking extract (CSE). Luminex assays and Western blots were employed to examine inflammatory status and pathological phenotypes of OFs. A specific reactive oxygen species (ROS) probe was used to evaluate oxidative stress levels. RNA-sequencing of CSE-treated OFs was used to analyze differentially expressed genes. Immunofluorescence and RNA-sequencing were used to examine the expression of receptor for advanced glycation end products (RAGE) signaling molecules in patients. Small interfering RNA sequences and a RAGE-specific inhibitor were employed to investigate the effects of RAGE blockade on cigarette smoking-related pathological phenotypes. To validate our findings , we generated an adenovirus-induced TED mouse model with exposure to cigarette smoke. Exposure to CSE resulted in an inflammatory phenotype of OFs together with higher levels of oxidative stress. OFs exposed to CSE presented susceptibility to transforming growth factor-β-induced myofibroblast differentiation, and 15-D-PGJ-induced adipocyte differentiation, indicating pro-fibrotic and pro-adipogenic phenotypes. RNA-sequencing of CSE-treated OFs revealed upregulation of RAGE signaling molecules. TED patients with smoking history also exhibited higher levels of RAGE signaling, both in the orbit and peripheral blood, compared with non-smoking patients. Enhancement of inflammatory status was associated with activation of the ROS-nuclear factor-kappa B pathway downstream of RAGE. RAGE gene interference or administration of RAGE inhibitor effectively mitigated cigarette smoking-related pathological changes in OFs. Disrupting RAGE signaling in TED mice efficiently ameliorated smoking-induced disease progression . Cigarette smoking-relevant TED progression was linked with RAGE signaling activation, leading to the exacerbation of orbital inflammation and tissue-remodeling, including fibrosis and adipogenesis. Our findings demonstrate that cigarette smoke exposure affects the biological characteristics of TED-derived OFs and supports RAGE as a promising therapeutic target for the management of patients with TED and smoking habits.
甲状腺眼病（TED）是一种威胁视力的自身免疫性疾病，吸烟是关键危险因素之一。吸烟会影响TED的严重程度以及患者对药物的反应。然而，吸烟在TED中的潜在致病机制仍不清楚。从TED患者和非TED对照中提取眼眶成纤维细胞（OFs），并用香烟烟雾提取物（CSE）处理。采用Luminex分析和蛋白质免疫印迹法检测OFs的炎症状态和病理表型。使用特异性活性氧（ROS）探针评估氧化应激水平。对经CSE处理的OFs进行RNA测序以分析差异表达基因。采用免疫荧光和RNA测序检测患者晚期糖基化终末产物受体（RAGE）信号分子的表达。使用小干扰RNA序列和RAGE特异性抑制剂研究RAGE阻断对吸烟相关病理表型的影响。为了验证我们的发现，我们构建了暴露于香烟烟雾的腺病毒诱导的TED小鼠模型。暴露于CSE会导致OFs出现炎症表型以及更高水平的氧化应激。暴露于CSE的OFs对转化生长因子-β诱导的肌成纤维细胞分化和15-D-前列腺素J2诱导的脂肪细胞分化敏感，表明其具有促纤维化和促脂肪生成表型。对经CSE处理的OFs进行RNA测序显示RAGE信号分子上调。与不吸烟患者相比，有吸烟史的TED患者在眼眶和外周血中也表现出更高水平的RAGE信号。炎症状态的增强与RAGE下游的ROS-核因子-κB通路激活有关。RAGE基因干扰或给予RAGE抑制剂可有效减轻OFs中与吸烟相关的病理变化。破坏TED小鼠中的RAGE信号可有效改善吸烟诱导的疾病进展。与吸烟相关的TED进展与RAGE信号激活有关，导致眼眶炎症和组织重塑加剧，包括纤维化和脂肪生成。我们的研究结果表明，接触香烟烟雾会影响TED来源的OFs的生物学特性，并支持RAGE作为治疗有吸烟习惯的TED患者的有前景的治疗靶点。
