Kirindage Kirinde Gedara Isuru Sandanuwan, Jayasinghe Arachchige Maheshika Kumari, Han Eui-Jeong, Jee Youngheun, Kim Hyun-Jin, Do Sun Gil, Fernando Ilekuttige Priyan Shanura, Ahn Ginnae
Department of Food Technology and Nutrition, Chonnam National University, Yeosu 59626, Korea.
Department of Veterinary Medicine and Veterinary Medical Research Institute, Jeju National University, Jeju 63243, Korea.
Antioxidants (Basel). 2022 Jul 23;11(8):1429. doi: 10.3390/antiox11081429.
is a well-known edible brown alga that is widely abundant in the sea near China, Korea, and Japan and has a wide range of bioactive compounds. Fucosterol (FST), which is a renowned secondary metabolite in brown algae, was extracted from to 70% ethanol, isolated via high-performance liquid chromatography (HPLC), followed by the immiscible liquid-liquid separation, and its structure was confirmed by NMR spectroscopy. The present study was undertaken to investigate the effects of FST against oxidative stress, inflammation, and its mechanism of action in tumor necrosis factor (TNF)-α/ interferon (IFN)-γ-stimulated human dermal fibroblast (HDF). FST was biocompatible with HDF cells up to the 120 μM dosage. TNF-α/IFN-γ stimulation significantly decreased HDF viability by notably increasing reactive oxygen species (ROS) production. FST dose-dependently decreased the intracellular ROS production in HDFs. Western blot analysis confirmed a significant increment of nuclear factor erythroid 2-related factor 2 (Nrf2)/ heme oxygenase-1 (HO-1) involvement in FST-treated HDF cells. In addition, the downregulation of inflammatory mediators, molecules related to connective tissue degradation, and tissue inhibitors of metalloproteinases were identified. TNF-α/IFN-γ stimulation in HDF cells increased the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) mediators, and its phosphorylation was reduced with the treatment of FST in a dose-dependent manner. Results obtained from western blot analysis of the NF-κB nuclear translocation were supported by immunocytochemistry results. Collectively, the outcomes suggested that FST significantly upregulates the Nrf2/HO-1 signaling and regulates NF-κB/MAPK signaling pathways to minimize the inflammatory responses in TNF-α/IFN-γ-stimulated HDF cells.
是一种著名的可食用褐藻，在中国、韩国和日本附近海域广泛分布，含有多种生物活性化合物。岩藻甾醇（FST）是褐藻中一种著名的次生代谢产物，从至70%乙醇中提取，通过高效液相色谱（HPLC）分离，随后进行不混溶液-液分离，其结构通过核磁共振光谱确认。本研究旨在探讨FST对氧化应激、炎症的影响及其在肿瘤坏死因子（TNF）-α/干扰素（IFN）-γ刺激的人皮肤成纤维细胞（HDF）中的作用机制。FST在高达120 μM剂量下与HDF细胞具有生物相容性。TNF-α/IFN-γ刺激通过显著增加活性氧（ROS）产生显著降低HDF活力。FST剂量依赖性地降低HDFs中的细胞内ROS产生。蛋白质印迹分析证实FST处理的HDF细胞中核因子红细胞2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）的参与显著增加。此外，还鉴定了炎症介质、与结缔组织降解相关分子和金属蛋白酶组织抑制剂的下调。HDF细胞中的TNF-α/IFN-γ刺激增加了核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）介质的磷酸化，而FST处理以剂量依赖性方式降低了其磷酸化。蛋白质印迹分析NF-κB核转位的结果得到免疫细胞化学结果的支持。总体而言，结果表明FST显著上调Nrf2/HO-1信号并调节NF-κB/MAPK信号通路，以最小化TNF-α/IFN-γ刺激的HDF细胞中的炎症反应。
