Wang Bo-Wen, Zhu Ru, Jin Ying, Ouyang Xuan, Jiang Fa-Gang, Wang Xing-Hua
Department of Ophthalmology, Tongji Medical College, Union Hospital, Huazhong University of Science and Technology, Wuhan, China.
Aier Eye Hospital of Wuhan University, Wuhan, China.
Front Mol Biosci. 2025 Jul 23;12:1580062. doi: 10.3389/fmolb.2025.1580062. eCollection 2025.
Thyroid-associated ophthalmopathy (TAO) is an autoimmune orbital disorder characterized by pathological alterations including extraocular muscle fibrosis and orbital inflammation. Cerium oxide nanoparticles (CeO2-NPs, CNPs) are gaining popularity in ophthalmology due to their potential antifibrotic and anti-inflammatory properties. This study aims to investigate the inhibitory effects of CNPs on fibrosis and inflammation in TAO orbital fibroblasts (OFs) derived from TAO patients.
OFs obtained by primary culture of orbital adipose tissue from 8 TAO patients. Probing the safe action concentration of CNPs and Anisomycin using CCK8 and detecting intracellular reactive oxygen species (ROS) generation using ROS Assay kit. Wound-Healing Assay was used to examine the degree of fibrosis of OFs. The expression of Fibronectin, COL1A1, α-Smooth muscle action, Hyaluronan Synthase 2, c-Jun N-terminal Kinase (JNK) and pJNK were detected by the RT-PCR and WB, and Hyaluronic Acid secretion was detected by ELISA. Inflammatory factors Interleukin-6 (IL-6) and Tumor Necrosis Factor-α (TNFα) expression were detected by RT-PCR and ELISA.
CNPs below 100 μg/mL and Anisomycin below 5 μM did not affect OFs proliferation. CNPs inhibit intracellular ROS generation. CNPs inhibit OFs fibrosis and suppress the expression of fibrosis indicators. These antifibrotic effects were mediated by inhibition of JNK phosphorylation, and were reversible by a JNK agonist. Furthermore, CNPs reduce both mRNA levels and secretion of inflammatory factors, IL-6 and TNF-α.
CNPs demonstrated the ability to inhibit fibrosis in TAO OFs by reducing JNK phosphorylation, as well as dose-dependently suppressed ROS generation and inflammatory response in TAO OFs.
甲状腺相关眼病（TAO）是一种自身免疫性眼眶疾病，其特征为包括眼外肌纤维化和眼眶炎症在内的病理改变。氧化铈纳米颗粒（CeO2-NPs，CNPs）因其潜在的抗纤维化和抗炎特性而在眼科领域日益受到关注。本研究旨在探讨CNPs对TAO患者来源的TAO眼眶成纤维细胞（OFs）纤维化和炎症的抑制作用。
从8例TAO患者的眼眶脂肪组织中通过原代培养获得OFs。使用CCK8检测CNPs和茴香霉素的安全作用浓度，并使用ROS检测试剂盒检测细胞内活性氧（ROS）的产生。采用伤口愈合试验检测OFs的纤维化程度。通过RT-PCR和WB检测纤连蛋白、COL1A1、α-平滑肌肌动蛋白、透明质酸合酶2、c-Jun氨基末端激酶（JNK）和pJNK的表达，并通过ELISA检测透明质酸分泌。通过RT-PCR和ELISA检测炎性因子白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNFα）的表达。
低于100μg/mL的CNPs和低于5μM的茴香霉素不影响OFs的增殖。CNPs抑制细胞内ROS的产生。CNPs抑制OFs纤维化并抑制纤维化指标的表达。这些抗纤维化作用是通过抑制JNK磷酸化介导的，并且可被JNK激动剂逆转。此外，CNPs降低炎性因子IL-6和TNF-α的mRNA水平和分泌。
CNPs通过降低JNK磷酸化表现出抑制TAO OFs纤维化的能力，并剂量依赖性地抑制TAO OFs中的ROS产生和炎症反应。
