Department of Food Science and Biotechnology, Graduate School, Kyungpook National University, Daegu 41566, Korea.
Food and Bio-Industry Research Institute, Inner Beauty/Anti-Aging Center, Kyungpook National University, Daegu 41566, Korea.
Biomolecules. 2020 Jan 29;10(2):199. doi: 10.3390/biom10020199.
As part of our continuous effort to find potential anti-inflammatory agents from endophytic fungi, a strain, isolated from the plant Roxb., was investigated. Cerevisterol (CRVS) was identified from endophytic fungi, a strain, and moreover exhibited anti-inflammatory activity. However, the underlying mode of action remains poorly understood. The aim of this study is to reveal the potential mechanisms of CRVS against inflammation on a molecular level in LPS-activated RAW 264.7 peritoneal macrophage cells. CRVS was isolated from and characterized based on spectral data analysis. The MTT assay was performed to measure cell viability in CRVS-treated macrophages. Anti-inflammatory activity was assessed by measurement of nitric oxide (NO) and prostaglandin E (PGE) levels, as well as the production of various cytokines, such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and -6 (IL-6) in LPS-stimulated macrophages. RT-PCR and immunoblotting analyses were done to examine the expression of various inflammatory response genes. A reporter gene assay was conducted to measure the level of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein-1 (AP-1) transactivation. CRVS suppresses the LPS-induced production of NO and PGE, which is a plausible mechanism for this effect is by reducing the expression of iNOS and COX-2. CRVS also decreases the expression of pro-inflammatory cytokines, such as TNF-α, IL-6, and IL-1β. CRVS halted the nuclear translocation of NF-κB by blocking the phosphorylation of inhibitory protein κBα (IκBα) and suppressing NF-κB transactivation. The mitogen-activated protein kinases (MAPK) signaling pathways are also suppressed. CRVS treatment also inhibited the transactivation of AP-1 and the phosphorylation of c-Fos. Furthermore, CRVS could induce the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) by down-regulating Kelch-like ECH-associated protein 1 (Keap-1) and up-regulating hemeoxygenases-1 (HO-1) expression. The results suggest that CRVS acts as a natural agent for treating inflammatory diseases by targeting an MAPK, NF-κB, AP-1, and Nrf2-mediated HO-1 signaling cascade.
从植物内生真菌中寻找潜在抗炎药物是我们持续努力的一部分。从植物内生真菌中分离出的一种菌株被鉴定为，其代谢产物麦角甾醇（CRVS）具有抗炎活性。然而，其作用机制尚不清楚。本研究旨在从分子水平上揭示 CRVS 对 LPS 激活的 RAW 264.7 腹腔巨噬细胞炎症的潜在作用机制。CRVS 从植物内生真菌中分离得到，并通过光谱数据分析进行了鉴定。MTT 法检测 CRVS 处理巨噬细胞的细胞活力。通过测量一氧化氮（NO）和前列腺素 E（PGE）的水平以及肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）等各种细胞因子的产生来评估抗炎活性，在 LPS 刺激的巨噬细胞中。RT-PCR 和免疫印迹分析用于检测各种炎症反应基因的表达。通过报告基因测定测量核因子κB 轻链增强子的活化 B 细胞（NF-κB）和激活蛋白-1（AP-1）的转激活水平。CRVS 抑制 LPS 诱导的 NO 和 PGE 的产生，这可能是通过降低诱导型一氧化氮合酶（iNOS）和环加氧酶-2（COX-2）的表达来实现的。CRVS 还降低了 TNF-α、IL-6 和 IL-1β 等促炎细胞因子的表达。CRVS 通过阻断抑制蛋白κBα（IκBα）的磷酸化和抑制 NF-κB 转激活来阻止 NF-κB 的核易位。丝裂原活化蛋白激酶（MAPK）信号通路也受到抑制。CRVS 处理还抑制了 AP-1 的转激活和 c-Fos 的磷酸化。此外，CRVS 通过下调 Kelch 样 ECH 相关蛋白 1（Keap-1）和上调血红素加氧酶-1（HO-1）的表达，诱导核因子红细胞 2 相关因子 2（Nrf2）的核易位。结果表明，CRVS 通过靶向 MAPK、NF-κB、AP-1 和 Nrf2 介导的 HO-1 信号级联，作为一种天然药物治疗炎症性疾病。
Zotero 插件
