title: Myrsine seguinii ethanolic extract and its active component quercetin inhibit macrophage activation and peritonitis induced by LPS by targeting to Syk/Src/IRAK-1.
abstract: BACKGROUND
  Myrsine seguinii H. LÉVEILLÉ (syn. Rapanea neriifolia) (Myrsinaceae) is a medicinal plants traditionally used in Myanmar to treat infectious and inflammatory diseases. Since none of reports have systematically demonstrated the anti-inflammatory activity of this plant, we aimed to mechanistically understand the regulatory roles of the plant in inflammatory responses using the ethanolic extract of Myrsine seguinii (Ms-EE).
  
  
  METHODS
  Activated macrophages and peritonitis symptoms induced by lipopolysaccharide (LPS) were employed. HPLC analysis was used to identify active components. To characterize direct target enzymes, kinase assay was established.
  
  
  RESULTS
  Ms-EE inhibited the production of nitric oxide (NO) and prostaglandin (PG)E2 in RAW264.7 cells and peritoneal macrophages stimulated by LPS. This extract suppressed the mRNA expression of the inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 genes by down-regulating the activation of nuclear factor (NF)-κB and activator protein (AP-1). Interestingly, it was found that Ms-EE can directly suppress the enzyme activities of Syk, Src, and interleukin-1 receptor-associated kinase-1 (IRAK-1). Similarly, orally administered Ms-EE inhibited the phosphorylation of Src and Syk in peritoneal exudate-derived cells prepared from peritonitis. Finally, HPLC analysis clearly demonstrated that quercetin is a major active component with suppressing activity on the release of inflammatory mediators (NO and PGE2), and the enzyme activities of Src, Syk, and IRAK-1.
  
  
  CONCLUSIONS
  Ms-EE containing quercetin negatively modulates macrophage-mediated in vitro inflammatory responses and LPS-induced peritonitis by blocking the Src/Syk/NF-κB and IRAK-1/AP-1 pathways, which contributes to its major ethnopharmacological use as an anti-inflammatory herbal medicine.
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Woo Seok
authors.initials: WS
authors.lastname: Yang
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Deok
authors.initials: D
authors.lastname: Jeong
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Young-Su
authors.initials: YS
authors.lastname: Yi
authors.affiliation: Division of Biological Resources Coordination, National Institute of Biological Resources, Incheon 404-708, Republic of Korea.
authors.forename: Byoung-Hee
authors.initials: BH
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, Kangwon National University, Chuncheon 200-701, Republic of Korea.
authors.forename: Tae Woong
authors.initials: TW
authors.lastname: Kim
authors.affiliation: Popa Mountain Park, Forest Department, Kyaukpadaung Township, Mandalay Division, Myanmar.
authors.forename: Khin Myo
authors.initials: KM
authors.lastname: Htwe
authors.affiliation: Department of Life Science, Hallym University, Chuncheon 200-702, Republic of Korea.
authors.forename: Young-Dong
authors.initials: YD
authors.lastname: Kim
authors.affiliation: College of Pharmacy, The Catholic University of Korea, Bucheon 420-743, Republic of Korea. Electronic address: kdyoon@catholic.ac.kr.
authors.forename: Kee Dong
authors.initials: KD
authors.lastname: Yoon
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Sungyoul
authors.initials: S
authors.lastname: Hong
authors.affiliation: Department of Forest Sciences, Seoul National University, Seoul 151-921, Republic of Korea.
authors.forename: Woo-Shin
authors.initials: WS
authors.lastname: Lee
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: jaecho@skku.edu.
authors.forename: Jae Youl
authors.initials: JY
authors.lastname: Cho
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D015232:Dinoprostone / Q000378:metabolism; D000431:Ethanol / Q000737:chemistry; D006801:Humans; D053592:Interleukin-1 Receptor-Associated Kinases / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008070:Lipopolysaccharides; D008262:Macrophage Activation / Q000187:drug effects; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000378:metabolism; D010538:Peritonitis / Q000188:drug therapy* / Q000378:metabolism; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology* / Q000627:therapeutic use*; D018515:Plant Leaves; D018547:Plant Stems; D029623:Primulaceae; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012997:Solvents / Q000737:chemistry; D000072377:Syk Kinase; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D010936:Plant Extracts; D012997:Solvents; D009569:Nitric Oxide; D000431:Ethanol; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C000606897:Syk protein, mouse; D019061:src-Family Kinases; C507768:IRAK1 protein, human; D053592:Interleukin-1 Receptor-Associated Kinases; D015232:Dinoprostone
keywords: (TNF)-α; 3,4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole; AP-1; ATF2; Akt; Anti-inflammatory effects; COX; CREB; EIA; ERK; IKK; IRAK-1; IκBα kinase; JNK; LPS; MAP kinase kinase; MAPK; MKK; MTT; Ms-EE; MyD88; Myrsine seguinii H. LÉVEILLÉ; NF-κB; NO; Nitric oxide; PGE(2); PI3K; Prostaglandin E(2); Quercetin; RT-PCR; Syk; TAK1; TGF-beta-activated kinase-1; TIR-domain-containing adapter-inducing interferon-β; TLR; TRIF; Tumour necrosis factor-α; activating transcription factor 2; activator protein-1; c-Jun N-terminal kinase; cAMP response element-binding; cyclooxygenase; enzyme immunoassay; ethanolic extract of Myrsine seguinii; extracellular signal-related kinase; iNOS; inducible NO synthase; interleukin-1 receptor-associated kinase-1; lipopolysaccharide; mitogen-activated protein kinase; myeloid differentiation primary response protein-88; nitric oxide; nuclear factor-κB; phosphoinositide 3-kinase; prostaglandin E(2); protein kinase B; reverse transcriptase-polymerase chain reaction; spleen tyrosine kinase; toll-like receptors (TLR); tumour necrosis factor