Src Is a Prime Target Inhibited by Celtis choseniana Methanol Extract in Its Anti‐Inflammatory Action

Abstract

Celtis choseniana is the traditional plant used at Korea as a herbal medicine to ameliorate inflammatory responses. Although Celtis choseniana has been traditionally used as a herbal medicine at Korea, no systemic research has been conducted on its anti-inflammatory activity. Therefore, the present study explored an anti-inflammatory effect and its underlying molecular mechanism using Celtis choseniana methanol extract (Cc-ME) in macrophage-mediated inflammatory responses. In vitro anti-inflammatory activity of Cc-ME was evaluated using RAW264.7 cells and peritoneal macrophages stimulated by lipopolysaccharide (LPS), pam3CSK4 (Pam3), or poly(I:C). In vivo anti-inflammatory activity of Cc-ME was investigated using acute inflammatory disease mouse models, such as LPS-induced peritonitis and HCl/EtOH-induced gastritis. The molecular mechanism of Cc-ME-mediated anti-inflammatory activity was examined by Western blot analysis and immunoprecipitation using whole cell and nuclear fraction prepared from the LPS-stimulated RAW264.7 cells and HEK293 cells. Cc-ME inhibited NO production and mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), and tumor necrosis factor-alpha (TNF-α) in the RAW264.7 cells and peritoneal macrophages induced by LPS, pam3, or poly(I:C) without cytotoxicity. High-performance liquid chromatography (HPLC) analysis showed that Cc-ME contained anti-inflammatory flavonoids quercetin, luteolin, and kaempferol. Among those, the content of luteolin, which showed an inhibitory effect on NO production, was highest. Cc-ME suppressed the NF-κB signaling pathway by targeting Src and interrupting molecular interactions between Src and p85, its downstream kinase. Moreover, Cc-ME ameliorated the morphological finding of peritonitis and gastritis in the mouse disease models. Therefore, these results suggest that Cc-ME exerted in vitro and in vivo anti-inflammatory activity in LPS-stimulated macrophages and mouse models of acute inflammatory diseases. This anti-inflammatory activity of Cc-ME was dominantly mediated by targeting Src in NF-κB signaling pathway during macrophage-mediated inflammatory responses.

Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties

“…To determine the cytotoxicity of Pc-EE, cells were treated with 25, 50, 100, or 200 μ g/mL Pc-EE in each experimental condition. To test the effects of Pc-EE on UVB- or H 2 O 2 -induced toxicity in HaCaT cells, the cells were treated with either 30 mJ/cm 2 UVB or 50 mM H 2 O 2 and cultured in complete culture medium with 50, 100, or 200 μ g/mL Pc-EE for 24 h. Cell viability was determined using a conventional MTT assay [20]. The cell viability was expressed as a percent using the following formula: cell viability (%) = [A0/A1] × 100, where A0 is the absorbance of the sample mean value and A1 is the absorbance of the normal group mean value.…”

Src/NF-κB-Targeted Anti-Inflammatory Effects of Potentilla glabra var. Mandshurica (Maxim.) Hand.-Mazz. Ethanol Extract

“…RAW264.7 cells (1 × 10 6 cells/mL) were pretreated with Pg-EE (150 µg/mL) for 30 min and subsequently, LPS (1 µg/mL) induction was processed for a designated time. HEK293T cells were transfected with a specific gene, incubated for 24 h, and then processed with Pg-EE for another 24 h. Whole cell lysates and stomach lysates of gastritis mice were prepared for western blot analysis, as previously described [33,34]. Western blot analyses using whole cell lysates and stomach lysates were conducted as previously described using antibodies specific for each target protein [33].…”

Trichosanthes tricuspidata Lour. Methanol Extract Exhibits Anti-Inflammatory Activity by Targeting Syk, Src, and IRAK1 Kinase Activity

“…For cell-based experiments, Tt-ME stock solution (200 μg/ml) was prepared in dimethyl sulfoxide (DMSO). For animal experiments, Tt-ME (200 mg/ml) was dissolved in 0.5% sodium carboxymethyl cellulose (sodium CMC), and drug treatment was performed as previously described [16,17].…”