Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea.
Department of Biochemistry and Molecular Biology, Yonsei University College of Medicine, Seoul, Korea.
Endocr J. 2020 Apr 28;67(4):439-447. doi: 10.1507/endocrj.EJ19-0521. Epub 2020 Jan 16.
Oxidative stress and adipogenesis play key roles in the pathogenesis of Graves' orbitopathy (GO). In this study, the therapeutic effects of caffeine on the reduction of oxidative stress and adipogenesis were evaluated in primary cultured GO orbital fibroblasts in vitro. Orbital fibroblasts were cultured from orbital connective tissues obtained from individuals with GO. Intracellular reactive oxygen species (ROS) levels induced by hydrogen peroxide or cigarette smoke extract and the expression of anti-oxidative enzymes were measured after caffeine treatment. After adipogenic differentiation and caffeine treatment, cells were stained with Oil Red O and the levels of peroxisome proliferator activator γ (PPARγ), C/EBPα, and C/EBPβ were determined by western blot analysis. Hydrogen peroxide and cigarette smoke extract increased the levels of intracellular ROS and anti-oxidative enzymes, which decreased in a dose-dependent manner upon pretreatment with caffeine in GO orbital fibroblasts. Oil Red-O staining results revealed a decrease in lipid droplets; furthermore, PPARγ, C/EBPα, and C/EBPβ protein expression levels were inhibited upon treatment with caffeine during adipocyte differentiation. In conclusion, caffeine decreased oxidative stress and adipogenesis in GO orbital fibroblasts in vitro. These findings may contribute to the development of new types of caffeine-containing pharmacological agents for use in the management of GO.
氧化应激和脂肪生成在格雷夫斯眼病 (GO) 的发病机制中起着关键作用。在这项研究中，评估了咖啡因对体外原代培养 GO 眼眶成纤维细胞氧化应激和脂肪生成减少的治疗作用。眼眶成纤维细胞从 GO 患者的眼眶结缔组织中培养而来。用咖啡因处理后，测量过氧化氢或香烟烟雾提取物诱导的细胞内活性氧 (ROS) 水平和抗氧化酶的表达。在脂肪生成分化和咖啡因处理后，用油红 O 染色，并用 Western blot 分析测定过氧化物酶体增殖物激活受体 γ (PPARγ)、C/EBPα 和 C/EBPβ 的水平。过氧化氢和香烟烟雾提取物增加了细胞内 ROS 和抗氧化酶的水平，而在 GO 眼眶成纤维细胞中用咖啡因预处理时，这些水平呈剂量依赖性降低。油红 O 染色结果显示脂质滴减少；此外，在脂肪细胞分化过程中用咖啡因处理时，PPARγ、C/EBPα 和 C/EBPβ 蛋白表达水平受到抑制。总之，咖啡因可降低 GO 眼眶成纤维细胞的氧化应激和脂肪生成。这些发现可能有助于开发新型含咖啡因的药物制剂，用于 GO 的治疗。
