Syk/NF-κB-targeted anti-inflammatory activity of Melicope accedens (Blume) T.G. Hartley methanol extract. (10th May 2021)

• Record Type: Journal Article
• Authors: Kim, Jin Kyeong; Choi, Eunju; Hong, Yo Han; Kim, Haeyeop; Jang, Young-Jin; Lee, Jong Sub; Choung, Eui Su; Woo, Byoung Young; Hong, Yong Deog; Lee, Sarah; Lee, Byoung-Hee; Bach, Tran The; Kim, Ji Hye; Kim, Jong-Hoon; Cho, Jae Youl
• Is Part Of: Journal of ethnopharmacology. Volume 271(2021)
• Publication Date: 2021-05-10
• Subjects: Melicope accedens (Blume) T.G. hartley -- Flavonoids -- Quercetin -- Anti-Inflammation -- NF-κB -- Syk -- Gastritis; Ma-ME Melicope accedens (Blume) T.G. Hartley methanol extract -- LPS lipopolysaccharide -- NO nitric oxide -- RT-PCR reverse transcriptase-polymerase chain reaction -- HPLC high performance liquid chromatography -- iNOS inducible nitric oxide synthase -- COX-2 cyclooxygenase-2 -- IL interleukin -- NF-κB nuclear factor-kappa B -- IκBα inhibitor of nuclear factor-kappa B α -- TLR Toll-like receptor -- RPMI 1640 Roswell Park Memorial Institute 1640 -- DMEM Dulbecco's modified Eagle's medium -- FBS fetal bovine serum -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide -- PEI polyethylenimine; Ethnopharmacology -- Periodicals; Pharmacognosy -- Periodicals; Herbs -- Periodicals; Herbs -- Periodicals; Pharmacognosy -- Periodicals; Pharmacognosie -- Périodiques; Herbes -- Périodiques; 615.1
• DOI: 10.1016/j.jep.2021.113887
• Languages: English
• ISSNs: 0378-8741

Abstract

Abstract: Ethnopharmacological relevance: Melicope accedens (Blume) Thomas G. Hartley is a plant included in the family Rutaceae and genus Melicope . It is a native plant from Vietnam that has been used for ethnopharmacology. In Indonesia and Malaysia, the leaves of M. accedens are applied externally to decrease fever. Aim of the study: The molecular mechanisms of the anti-inflammatory properties of M. accedens are not yet understood. Therefore, we examined those mechanisms using a methanol extract of M. accedens (Ma-ME) and determined the target molecule in macrophages. Materials and methods: We evaluated the anti-inflammatory effects of Ma-ME in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and in an HCl/EtOH-triggered gastritis model in mice. To investigate the anti-inflammatory activity, we performed a nitric oxide (NO) production assay and ELISA assay for prostaglandin E2 (PGE2 ). RT-PCR, luciferase gene reporter assays, western blotting analyses, and a cellular thermal shift assay (CETSA) were conducted to identify the mechanism and target molecule of Ma-ME. The phytochemical composition of Ma-ME was analyzed by HPLC and LC-MS/MS. Results: Ma-ME suppressed the production of NO and PGE2 and the mRNA expression of proinflammatory genes (iNOS, IL-1β, and COX-2) in LPS-stimulated RAW264.7 cells without cytotoxicity. Ma-ME inhibited NF-κB activation by suppressing signaling molecules such as IκBα, Akt, Src, and Syk. Moreover, the CETSA assay revealed that Ma-ME binds to Syk, the most upstream molecule in the NF-κB signal pathway. Oral administration of Ma-ME not only alleviated inflammatory lesions, but also reduced the gene expression of IL-1β and p-Syk in mice with HCl/EtOH-induced gastritis. HPLC and LC-MS/MS analyses confirmed that Ma-ME contains various anti-inflammatory flavonoids, including quercetin, daidzein, and nevadensin. Conclusions: Ma-ME exhibited anti-inflammatory activities in vitro and in vivo by targeting Syk in the NF-κB signaling pathway. Therefore, we propose that Ma-ME could be used to treat inflammatory diseases such as gastritis. Graphical abstract: Image 1