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 proinflammatory 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). Conclusions 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.

Keywords

• inflammation
• mrna
• raw 264.7 cell line