Department of Ophthalmology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, Guangdong 519000, PR China.
Department of Pathology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, Guangdong 519000, PR China.
Exp Eye Res. 2022 Mar;216:108946. doi: 10.1016/j.exer.2022.108946. Epub 2022 Jan 14.
Chemokines and adhesion molecules are major inflammatory mediators of chronic and recurrent vernal keratoconjunctivitis (VKC). Sulforaphane (SFN) is a natural plant extract that is known to have anti-inflammatory and antioxidant properties. SFN is demonstrated to be effective against a variety of human diseases. The current investigation examines the effects and the molecular mechanisms of SFN on cytokine-induced human corneal fibroblasts (HCFs) expression of adhesion molecules and chemokines. HCFs were exposed to both interleukin (IL)-4 and tumor necrosis factor (TNF)-α in the absence or presence of SFN treatment. The levels of thymus- and activation-regulated chemokine (TARC) and eotaxin-1 in culture supernatants were evaluated using enzyme-linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction analysis (RT-PCR) enabled quantification of mRNA levels of vascular cell adhesion molecule (VCAM)-1, eotaxin-1, and TARC along with cytokine receptors. An immunoblotting assay was used to evaluate the activities of VCAM-1, nuclear factor-kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), signal transducer and activator of transcription factor (STAT)6 pathways, along with the expression of the cytokine receptors including IL-4 receptor (R)α, IL-13Rα1, TNFRI, as well as TNFRII. SFN inhibited TARC and eotaxin-1 release in HCFs stimulated by TNF-α and IL-4 in a manner dependent on dose and time. SFN suppressed transcriptions of TARC, eotaxin-1, and VCAM-1. Furthermore, the mRNA and protein expression levels of IL-4Rα, TNFRI, and TNFRII were also attenuated by SFN exposure, however, those of IL-13Rα1 remained unaffected. In addition, SFN downregulated the expression of VCAM-1 and the phosphorylation of MAPKs, IκBα, and STAT6. These results suggest that SFN inhibited cytokine-stimulated TARC, eotaxin-1 secretion as well as VCAM-1 expression in HCFs, with these effects likely occurring as a result of cytokine receptor inhibition and attenuation of MAPK, NF-κB, and STAT6 signaling. SFN may therefore have therapeutic potential in VKC treatment.
趋化因子和黏附分子是慢性和复发性春季角结膜炎 (VKC) 的主要炎症介质。萝卜硫素 (SFN) 是一种天然植物提取物，具有抗炎和抗氧化特性。SFN 已被证明对多种人类疾病有效。目前的研究检查了 SFN 对细胞因子诱导的人角膜成纤维细胞 (HCF) 黏附分子和趋化因子表达的影响及其分子机制。在存在或不存在 SFN 处理的情况下，将 HCF 暴露于白细胞介素 (IL)-4 和肿瘤坏死因子 (TNF)-α 中。使用酶联免疫吸附测定 (ELISA) 评估培养上清液中胸腺激活调节趋化因子 (TARC) 和嗜酸性粒细胞趋化因子-1(eotaxin-1) 的水平。逆转录-聚合酶链反应分析 (RT-PCR) 使血管细胞黏附分子 (VCAM)-1、eotaxin-1 和 TARC 的 mRNA 水平以及细胞因子受体的定量成为可能。免疫印迹分析用于评估 VCAM-1、核因子-κB (NF-κB)、丝裂原激活蛋白激酶 (MAPKs)、信号转导和转录激活因子 (STAT)6 途径的活性，以及细胞因子受体的表达，包括白细胞介素 (IL)-4 受体 (R)α、白细胞介素-13 受体 α1、TNFRI 以及 TNFRII。SFN 以剂量和时间依赖的方式抑制 TNF-α 和 IL-4 刺激的 HCF 中 TARC 和 eotaxin-1 的释放。SFN 抑制 TARC、eotaxin-1 和 VCAM-1 的转录。此外，SFN 暴露还减弱了 IL-4Rα、TNFRI 和 TNFRII 的 mRNA 和蛋白表达水平，但 IL-13Rα1 不受影响。此外，SFN 下调了 VCAM-1 的表达和 MAPKs、IκBα 和 STAT6 的磷酸化。这些结果表明，SFN 抑制了细胞因子刺激的 HCF 中 TARC、eotaxin-1 的分泌以及 VCAM-1 的表达，这些作用可能是由于细胞因子受体抑制以及 MAPK、NF-κB 和 STAT6 信号转导的衰减所致。SFN 因此可能具有治疗 VKC 的潜力。
