Anti-inflammatory Effects of Complex Extract including Eucommia ulmoides in LPS-induced RAW 264.7 Cells

Abstract

Background: The purpose of this study was to investigate the anti-inflammatory response of lipopolysaccharide (LPS) activated macrophages (RAW 264.7 murine cell line) to JCE003 which is an extract including Eucommia ulmoides, Juglans regia, Eleutherococcus senticosus, and Zingiber officinale. Methods: An MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed to analyze the survival rate of RAW 264.7 cells. The production of nitric oxide and pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β, IL-6) in LPS-induced RAW 264.7 cells was measured by enzyme-linked immunosorbent assay. mRNA expression levels of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β, and IL-6) were analyzed by quantitative polymerase chain reaction analysis. Results: Exposure of LPS-activated RAW 264.7 cells to JCE003 was not cytotoxic up to 400 μg/mL, but cell survival was statistically significantly decreased at 800 μg/mL (p < 0.001). Nitric oxide production was not markedly lowered in LPS-activated RAW 264.7 cells by exposure to JCE003 (10, 50, 100, 200, 400, 800 μl/mL) compared with the Control group. In addition, JCE003 reduced the production of TNF-α in LPS-induced RAW 264.7 cells at 400 μg/mL (p < 0.05), but IFN-γ and TNF-α mRNA expression in LPS-induced RAW 264.7 cells was decreased at 100, 200, and 400 μg/mL JCE003 (p < 0.01). Conclusion: These results suggest that JCE003 inhibited the expression and production of pro-inflammatory cytokines in LPS-activated RAW 264.7 cells. The findings of this study provide basic data for the development of new Korean medicine anti-inflammatory drugs.