Lee Chae Eun, Choi Soo Hyun, Yoon Jin Sook
Yonsei University College of Medicine, Seoul, Korea.
Institute of Vision Research, Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Korea.
Korean J Ophthalmol. 2020 Jun;34(3):192-202. doi: 10.3341/kjo.2020.0002.
Chemokines are involved in the pathogenesis of various autoimmune diseases, including Graves' orbitopathy (GO), but comprehensive analyses of the dynamics of these cytokines and their receptors in such diseases remain lacking. In this study, we investigated the expressions of chemokines and their receptors during adipogenesis and inflammation in primary cultured orbital fibroblasts from patients with GO.
The messenger RNA (mRNA) expression levels of chemokines were compared between GO (n = 6) and non-GO (n = 5) orbital tissues by real-time polymerase chain reaction. After adipogenesis was induced in primary cultured orbital fibroblasts from patients with GO (n =5) and following stimulation with interleukin (IL)-1β and tumor necrosis factor (TNF)-α, the mRNA expression levels of chemokines and their receptors were analyzed.
Chemokines were significantly downregulated in GO orbital tissues compared to non-GO orbital tissues ( < 0.05). Adipogenesis resulted in a strong increase in mRNA expression levels of chemokines and their receptors at an early stage (day 1); however, expression levels started to decrease thereafter and, eventually, decreased to below basal levels at the end of adipogenesis (day 10). Following stimulation with IL-1β and TNF-α, the mRNA expression levels of chemokines and their receptors increased, showing different responses to various proinflammatory cytokines.
Chemokines were strongly upregulated in the early phase of adipogenesis before decreasing continuously until the end of adipogenesis. Also, overt mature GO tissues showed reduced mRNA expression of chemokines compared to controls, which might indicate the existence of a shorter window for effective medical inflammatory treatment. The heightened levels of chemokines and their receptors observed after stimulation with IL-1β and TNF-α suggest a crucial role of proinflammatory cytokines in the pathogenesis of GO and, further, support the idea that chemokines could be used as biomarkers of GO activity.
趋化因子参与包括格雷夫斯眼眶病（GO）在内的多种自身免疫性疾病的发病机制，但对这些细胞因子及其受体在这类疾病中的动态变化进行全面分析的研究仍很缺乏。在本研究中，我们调查了GO患者原代培养的眼眶成纤维细胞在脂肪生成和炎症过程中趋化因子及其受体的表达情况。
通过实时聚合酶链反应比较GO组（n = 6）和非GO组（n = 5）眼眶组织中趋化因子的信使核糖核酸（mRNA）表达水平。在诱导GO患者（n = 5）原代培养的眼眶成纤维细胞发生脂肪生成后，以及在用白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α刺激后，分析趋化因子及其受体的mRNA表达水平。
与非GO眼眶组织相比，GO眼眶组织中的趋化因子显著下调（<0.05）。脂肪生成在早期阶段（第1天）导致趋化因子及其受体的mRNA表达水平大幅增加；然而，此后表达水平开始下降，最终在脂肪生成结束时（第10天）降至基础水平以下。在用IL-1β和TNF-α刺激后，趋化因子及其受体的mRNA表达水平升高，显示出对各种促炎细胞因子的不同反应。
趋化因子在脂肪生成的早期阶段强烈上调，然后持续下降直至脂肪生成结束。此外，与对照组相比，明显成熟的GO组织中趋化因子的mRNA表达降低，这可能表明有效药物抗炎治疗的窗口期较短。在用IL-1β和TNF-α刺激后观察到的趋化因子及其受体水平升高表明促炎细胞因子在GO发病机制中起关键作用，进一步支持了趋化因子可作为GO活动生物标志物的观点。
