Nitidine chloride inhibits LPS-induced inflammatory cytokines production via MAPK and NF-kappaB pathway in RAW 264.7 cells
Nitidine chloride inhibits LPS-induced inflammatory cytokines production via MAPK and NF-kappaB pathway in RAW 264.7 cells
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氯化两面针碱通过 MAPK 和 NF-kappaB 途径抑制 RAW 264.7 细胞中 LPS 诱导的炎症细胞因子的产生
DOI：
10.1016/j.jep.2012.08.041
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发表时间：
2012-10-31
影响因子：
5.4
通讯作者：
Du, Bing
中科院分区：
文献类型：
--
作者：
Wang, Ziqiang;Jiang, Wei;Du, Bing
民族药理学相关性：两面针（Zanthoxylum nitidium (Roxb.) DC.）长期以来一直被用作治疗风湿性关节炎和牙周炎等炎症性疾病的传统草药。然而，氯化两面针碱的抗炎机制尚未完全阐明。
研究目的：确定从两面针根部提取的五环生物碱氯化两面针碱（Nitidine chloride，NTD）在小鼠巨噬细胞中的抗炎作用及其机制。
材料和方法：以脂多糖（LPS）刺激的Raw 264.7巨噬细胞作为体外模型，研究NTD的抗炎特性。通过实时逆转录聚合酶链反应（real - time RT - PCR）和酶联免疫吸附测定（ELISA）评估促炎细胞因子。此外，使用特异性抗体通过蛋白质印迹（Western blot）和免疫荧光染色分析细胞内信号通路。
结果：NTD在RNA和蛋白质水平上均显著降低了肿瘤坏死因子α（TNF - α）、白细胞介素 - 1β（IL - 1β）和IL - 6等促炎细胞因子的产生。此外，NTD以剂量依赖的方式显著抑制LPS处理的RAW 264.7细胞中核因子κB（NF - κB）的转录活性以及丝裂原活化蛋白激酶（MAPKs）的磷酸化。这些结果表明，NTD通过抑制RAW 264.7细胞中TNF - α、IL - 1β和IL - 6的产生以及减少NF - κB和MAPK信号通路来发挥抗炎特性。
结论：这些结果表明，NTD通过抑制RAW 264.7细胞中TNF - α、IL - 1β和IL - 6的产生以及减少NF - κB和MAPK信号通路来发挥抗炎特性。氯化两面针碱通过抑制MAPK的磷酸化和p65的易位来抑制LPS诱导的TNF - α、IL - 1β和IL - 6的产生。此外，这些结果揭示了NTD在炎症性疾病调节中的新作用。（C）2012爱思唯尔爱尔兰有限公司。保留所有权利。
Ethnopharmacological relevance: Zanthoxylum nitidium (Roxb.) DC. has long been used as a traditional herbal medicine for inflammatory diseases such as rheumatic arthritis and peridentitis. However, the anti-inflammatory mechanism of Nitidine chloride has not been fully elucidated.Aim of the study: To determine the anti-inflammatory effects and mechanism of Nitidine chloride (NTD), a pentacyclic alkaloid is isolated from the root of Z. nitidium, in murine macrophages.Materials and methods: Anti-inflammatory properties of NTD were investigated using lipopolysaccharide (LPS)-stimulated Raw 264.7 macrophages as in vitro model. The pro-inflammatory cytokines were evaluated by real-time RT-PCR and ELISA. Furthermore, intracellular signaling pathways were analyzed by Western blot and Immunofluorescence staining using specific antibodies.Results: NTD significantly reduced the production of pro-inflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and IL-6 in both RNA and protein level. Moreover, transcriptional activity of NF-kappa B as well as the phosphorylation of mitogen-activated protein kinases (MAPKs) in LPS-treated RAW 264.7 was significantly inhibited by NTD in a dose dependent manner. These results suggested that NTD exerts an anti-inflammatory property by inhibiting TNF-alpha, IL-1 beta, and IL-6 production in association with reduced NF-kappa B and MAPK signaling pathways in RAW 264.7 cells.Conclusion: These results suggested that NTD exerts an anti-inflammatory property by inhibiting TNF-alpha, IL-1 beta, and IL-6 production in association with reduced NF-kappa B and MAPK signaling pathways in RAW 264.7 cells. Nitidine chloride inhibits LPS-induced TNF alpha, IL-1 beta and IL-6 production via the suppression of phosphorylation of MAPK and the translocation of p65. In addition, these results revealed a novel role of NTD in regulation of inflammatory diseases. (C) 2012 Elsevier Ireland Ltd. All rights reserved.