Kim Na-Hyun, Lee Seon Min, Kim Yun Na, Jeon You-Jin, Heo Jeong-Doo, Jeong Eun Ju, Rho Jung-Rae
Gyeongnam Department of Environment & Toxicology, Korea Institute of Toxicology, 17 Jegok-gil, Munsan-eup 5834, Korea.
Department of Agronomy and Medicinal Plant Resources, Gyeongnam National University of Science and Technology, Jinju 52725, Korea.
Biomolecules. 2020 Oct 20;10(10):1463. doi: 10.3390/biom10101463.
is a tropical brown algae (seaweed) known to have anti-inflammatory properties. In this study, we analyzed extract (TOE) using liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) and nuclear magnetic resonance (NMR) and evaluated the in vivo efficacy of TOE against dextran sulfate sodium-induced chronic colitis in C57BL/6 mice. The bioactive fraction of TOE was administered orally daily for 6 weeks to mice under different treatments normal, colitis, and colitis + conventional drug (5-aminosalicylic acid, 5-ASA). Regarding clinical manifestation, the disease activity index and colon length of the colitis + TOE group were significantly reduced compared to those of the colitis group. The results of myeloperoxidase activity and histopathological examination showed similar results. Western blot analysis of colon tissues revealed that cyclooxygenase-2, tumor necrosis factor alpha (TNF-α), and phosphorylated signal transducer and activator of transcription-3 (p-STAT3) were significantly decreased in the colitis + 5-ASA group, whereas forkhead box P3 (FOXP3) was increased. qPCR results showed changes in T cell subsets; the administration of TOE upregulated regulatory T cell (Treg) expression, although T helper 17 cell (Th17) expression did not change significantly. Interestingly, the colitis + TOE group showed high levels of both Th1 and Th2 transcription factors, but the secreted cytokine interferon (IFN)-γ and interleukin (IL)-4 remained unchanged and somewhat reduced. Additionally, TNF-α gene expression was significantly reduced in the colitis + TOE group. IL-6 mRNA levels were also decreased, although not significantly. Four compounds were structurally elucidated using 1D- and 2D-NMR spectroscopy, and five compounds were fully identified or tentatively characterized using LC-QTOF-MS. In conclusion, TOE could alleviate chronic colitis via upregulation of Foxp3 Treg cells and production of the anti-inflammatory cytokine IL-10, which directly inhibits macrophages and pro-inflammatory cytokine synthesis, leading to reduced colitis.
是一种已知具有抗炎特性的热带褐藻（海藻）。在本研究中，我们使用液相色谱四极杆飞行时间质谱（LC-QTOF-MS）和核磁共振（NMR）分析了提取物（TOE），并评估了TOE对葡聚糖硫酸钠诱导的C57BL/6小鼠慢性结肠炎的体内疗效。将TOE的生物活性部分每日口服给药6周，用于不同处理的小鼠：正常、结肠炎和结肠炎 + 传统药物（5-氨基水杨酸，5-ASA）。关于临床表现，与结肠炎组相比，结肠炎 + TOE组的疾病活动指数和结肠长度显著降低。髓过氧化物酶活性和组织病理学检查结果显示出相似的结果。结肠组织的蛋白质印迹分析表明，结肠炎 + 5-ASA组中环氧合酶-2、肿瘤坏死因子α（TNF-α）和磷酸化信号转导和转录激活因子3（p-STAT3）显著降低，而叉头框P3（FOXP3）增加。定量聚合酶链反应（qPCR）结果显示T细胞亚群发生变化；TOE的给药上调了调节性T细胞（Treg）的表达，尽管辅助性T细胞17（Th17）细胞的表达没有显著变化。有趣的是，结肠炎 + TOE组显示出高水平的Th1和Th2转录因子，但分泌的细胞因子干扰素（IFN）-γ和白细胞介素（IL）-4保持不变且有所降低。此外，结肠炎 + TOE组中TNF-α基因表达显著降低。IL-6 mRNA水平也有所下降，尽管不显著。使用一维和二维NMR光谱对四种化合物进行了结构解析，并使用LC-QTOF-MS对五种化合物进行了完全鉴定或初步表征。总之，TOE可通过上调Foxp3 Treg细胞和产生抗炎细胞因子IL-10来减轻慢性结肠炎，IL-10可直接抑制巨噬细胞和促炎细胞因子的合成，从而减轻结肠炎。
