Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea.
Department of Endocrinology, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
Invest Ophthalmol Vis Sci. 2019 Oct 1;60(13):4129-4140. doi: 10.1167/iovs.19-27376.
We examined the therapeutic effect of nontoxic concentrations of curcumin, a plant polyphenol extracted from Curcuma longae, in primary cultures of orbital fibroblasts from Graves' orbitopathy (GO).
The effect of curcumin on interleukin (IL)-1β induced-proinflammatory cytokine production was determined using quantitative real-time PCR, enzyme-linked immunosorbent assay (ELISA), and Western blot analysis. Adipogenic differentiation was identified using Oil-Red O staining, and levels of peroxisome proliferator activator γ (PPARγ) and CCAAT-enhancer-binding proteins (C/EBP) α/β were determined by Western blot analyses. Antioxidant activity was measured using an oxidant-sensitive fluorescent probe to detect intracellular reactive oxygen species (ROS) generated in response to hydrogen peroxide (H2O2) and cigarette smoke extract (CSE).
Treatment with curcumin resulted in a dose- and time-dependent decrease in IL-1β-induced synthesis of inflammatory cytokines, including IL-6, IL-8, MCP-1, and ICAM-1 at both mRNA and protein levels. Decrease in lipid droplets and expression of PPARγ and c/EBPα/β were noted in fibroblasts treated with curcumin during adipose differentiation. CSE- or H2O2-induced ROS synthesis was significantly lower in curcumin-treated fibroblasts in comparison with the control. Curcumin significantly suppressed IL-1β-induced phosphorylated extracellular signal-regulated kinase, Akt, c-Jun NH(2)-terminal kinase, and nuclear factor κ-light-chain-enhancer of activated B cells, p65 proteins and stimulated β-catenin translocation into nucleus during adipogenesis.
Curcumin inhibits proinflammatory cytokine production, ROS synthesis, and adipogenesis in orbital fibroblasts of GO patients in vitro possibly related to multiple proinflammatory signaling molecules and β-catenin pathway. The results of the study support potential use of the curcumin in the treatment of GO.
我们研究了姜黄素（一种从姜黄中提取的植物多酚）的无毒浓度对格雷夫斯眼病（GO）眼眶成纤维细胞原代培养物中促炎细胞因子产生的治疗作用。
采用实时定量 PCR、酶联免疫吸附试验（ELISA）和 Western blot 分析检测姜黄素对白细胞介素（IL）-1β诱导的促炎细胞因子产生的影响。用油红 O 染色鉴定脂肪生成分化，用 Western blot 分析测定过氧化物酶体增殖物激活受体 γ（PPARγ）和 CCAAT 增强子结合蛋白（C/EBP）α/β的水平。用氧化敏感荧光探针检测过氧化氢（H2O2）和香烟烟雾提取物（CSE）刺激后细胞内活性氧（ROS）的产生来测定抗氧化活性。
姜黄素处理呈剂量和时间依赖性降低 IL-1β诱导的炎症细胞因子（包括 IL-6、IL-8、MCP-1 和 ICAM-1）的合成，mRNA 和蛋白水平均如此。在脂肪分化过程中，用姜黄素处理的成纤维细胞中可见脂滴减少，PPARγ 和 c/EBPα/β 的表达减少。与对照组相比，CSE 或 H2O2 诱导的 ROS 合成在姜黄素处理的成纤维细胞中明显降低。姜黄素显著抑制 IL-1β诱导的磷酸化细胞外信号调节激酶、Akt、c-Jun N-末端激酶和核因子 κB 轻链增强子的激活 B 细胞，p65 蛋白和刺激β-连环蛋白在脂肪生成过程中转位入核。
姜黄素抑制 GO 患者眼眶成纤维细胞中促炎细胞因子的产生、ROS 合成和脂肪生成，可能与多种促炎信号分子和β-连环蛋白途径有关。研究结果支持姜黄素在 GO 治疗中的潜在应用。
