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

Ethnopharmacological relevance: 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).

Materials and 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.

Conclusion: 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.

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.