Department of Integrative Biotechnology, Sungkyunkwan University, Suwon, 16419, Republic of Korea.
School of Systems Biomedical Science, Soongsil University, Seoul, 06978, Republic of Korea.
J Ethnopharmacol. 2019 Jun 28;238:111848. doi: 10.1016/j.jep.2019.111848. Epub 2019 Apr 2.
Canarium subulatum Guillaumin is an herbal medicinal plant native to Southeast Asia. Ethnopharmacological evidence suggests that plants of the genus Canarium cure a variety of inflammatory diseases.
The pharmacological mechanisms of C. subulatum Guillaumin remain poorly understood. In this study, we investigate inflammatory mechanisms and target molecules using C. subulatum Guillaumin methanol extract (Cs-ME) in inflammatory reactions managed by macrophages.
To identify the anti-inflammatory activities of Cs-ME, lipopolysaccharide (LPS)-stimulated macrophages and a murine HCl/EtOH-induced gastritis model were chosen. The luciferase reporter gene assay, Western blot analysis, overexpression strategy, and the cellular thermal shift assay (CETSA) were employed to investigate the molecular mechanisms and target enzymes of Cs-ME. The active ingredients of this extract were also determined by HPLC.
Released levels of nitric oxide (NO) and mRNA expression levels of iNOS and IL-6 were downregulated by Cs-ME without exhibiting cytotoxicity. This extract inhibited MyD88-induced promoter activity and the nuclear translocation of nuclear factor (NF)-κB. Moreover, we found that Cs-ME reduced the phosphorylation of NF-κB upstream signaling molecules including IκBα, IKKα/β, Src, and Syk in LPS-stimulated macrophage-like RAW264.7 cells. The results of Western blot and CETSA confirmed that Src and Syk are anti-inflammatory targets of Cs-ME. In addition, orally injected Cs-ME alleviated HCl/EtOH-induced gastric ulcers in mice. HPLC analysis indicated that quercetin, luteolin, and kaempferol are major active components of this extract with anti-inflammatory activity.
Cs-ME exhibits anti-inflammatory effects in vitro and in vivo by targeting Src and Syk in the NF-κB signaling pathway. Consequently, Cs-ME could be developed as an anti-inflammatory herbal medicine.
药用植物菓榄属植物治疗多种炎症性疾病的民族药理学证据。
药用植物菓榄属植物的药理机制仍知之甚少。本研究采用脂多糖（LPS）刺激的巨噬细胞和小鼠 HCl/EtOH 诱导的胃炎模型，研究菓榄属植物甲醇提取物（Cs-ME）在巨噬细胞炎症反应中的炎症机制和靶分子。
为了确定 Cs-ME 的抗炎活性，选择 LPS 刺激的巨噬细胞和小鼠 HCl/EtOH 诱导的胃炎模型。采用荧光素酶报告基因分析、Western blot 分析、过表达策略和细胞热转移分析（CETSA）研究 Cs-ME 的分子机制和靶酶。还通过 HPLC 确定了该提取物的活性成分。
Cs-ME 下调 LPS 诱导的巨噬细胞一氧化氮（NO）释放水平和诱导型一氧化氮合酶（iNOS）和白细胞介素 6（IL-6）的 mRNA 表达，且无细胞毒性。该提取物抑制 MyD88 诱导的启动子活性和核因子（NF）-κB 的核易位。此外，我们发现 Cs-ME 降低了 LPS 刺激的巨噬细胞样 RAW264.7 细胞中 NF-κB 上游信号分子包括 IκBα、IKKα/β、Src 和 Syk 的磷酸化。Western blot 和 CETSA 的结果证实 Src 和 Syk 是 Cs-ME 的抗炎靶点。此外，口服 Cs-ME 可减轻小鼠 HCl/EtOH 诱导的胃溃疡。HPLC 分析表明，槲皮素、木犀草素和山奈酚是该提取物的主要活性成分，具有抗炎活性。
Cs-ME 通过靶向 NF-κB 信号通路中的 Src 和 Syk，在体外和体内均表现出抗炎作用。因此，Cs-ME 可开发为一种抗炎草药。
