Abstract
Purpose: To investigate the antioxidant and anti-inflammatory effects of Ulmus davidiana extract (UDE) in lipopolysaccharide (LPS)-stimulated BV-2 cells. Methods: Antioxidant activity was measured using 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay. Cell viability was evaluated using 3 (4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) assay. BV-2 cells were stimulated with LPS to study protein expression and production of inflammatory mediators, and determined by Western blot analysis. Results: UDE significantly inhibited DPPH-generated free radicals showing maximum inhibition at 40 μg/mL (p < 0.001). UDE alone did not exhibit any signs of cytotoxicity towards BV-2 cells up to 100 μg/mL concentration. The LPS induced increase in the production of nitric oxide was concentration dependently suppressed with half-maximal concentration (IC 50 ) of 67.4 μg/ mL of UDE (p < 0.05 at 10 μg/mL, p < 0.01 at 20 μg/mL and p < 0.001 at 40 μg/mL, respectively). UDE also inhibited dosedependently the LPS-induced increase in inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expressions with IC 50 of 52.3 ug/ mL. Furthermore, the production of pro inflammatory cytokines, via tumor necrosis factor-α by LPS-stimulation in BV2 murine cells was inhibited dosedependently with IC 50 of 85.1 ug/ mL by UDE pretreatment. Mechanistic studies revealed that UDE acts by regulation of nuclear factor kappa-B signaling pathway in LPS-stimulated BV-2 cells. Conclusion: This study shows, for the first time, that UDE possesses antioxidant and anti-inflammatory effects and can be developed as a potential therapeutic agent for ameliorating macrophage-mediated inflammation. Keywords: Ulmus davidiana , Anti-oxidant, Anti-inflammatory, iNOS, NF-κB, MCP-1, ERK
Highlights
Ulmus davidiana var. japonica (UDE), from the family of Ulmaceae, is widely distributed in Korea, Japan, and China
Its proven medicinal potency merits an elucidation of the mechanisms by which it works and this study will focus on the molecular activities of Ulmus davidiana extract (UDE) on BV2 murine microglial cells
Previous reports demonstrate that suppressive agents of the inflammatory mediator concentrations in activated microglia can attenuate the severity of neuro-degenerative diseases [6]
Summary
Ulmus davidiana var. japonica (UDE), from the family of Ulmaceae, is widely distributed in Korea, Japan, and China. Cells were pre-treated with the indicated concentrations of UDE for 1 h before the addition of LPS (1 μg/mL, Sigma-Aldrich, St. Louis, MO, USA) in serum free DMEM. Cells were pretreated with the indicated concentrations of UDE for 1 h before the addition of LPS (1 μg/mL, Sigma-Aldrich, St. Louis, MO, USA) in serum free RPMI1640. UDE exhibited significant DPPH radical scavenging activity in a dosedependent manner, showing a maximum effect at 100 μg/mL of concentration (p < 0.001).
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