Study on Anti-inflammatory Activity and Mechanism of Compound 2-6-H-5-B from Eurotium Cristatum

OBJECTIVE To study the effects and mechanisms of anti-inflammatory of benzaldehyde derivatives (2-6-H-5-B) derived from Eurotium cristatum in lipopolysaccharide stimulated RAW264.7 cells. METHODS RAW264.7 cells were pretreated with benzaldehyde derivative(compound 2-6-H-5-B)(25, 50, 100 μmol·L^-1) and postive control drug dexamethasone(0.5 μmol·L^-1) for 1 h, then cells were treated with 100 ng·mL^-1 lipopolysaccharide for 24 h except control group. The nitrite concentration in the culture medium was measured as an indicator of NO production by the Griess reagent according to the manufacturer’s protocol. The expression levels of TNF-α, IL-6 and MCP-1 in the cultured supernatant were detected by ELISA kits, and the protein expressions of AMPK, NF-κB and MAPK signaling pathways were analyzed by Western blotting. RESULTS The expression of pro-inflammatory cytokines, including TNF-α(P<0.01), IL-6(P<0.01) and MCP-1(P<0.01), as well as the production of NO(P<0.01 or P<0.05) were inhibited by compound 2-6-H-5-B. Further mechanism studies showed that 2-6-H-5-B failed to inhibit the activation of NF-κB and MAPK signaling pathways, but increased the expression of p-AMPK significantly. CONCLUSION The 2-6-H-5-B is a potential anti-inflammatory drug, and its anti-inflammatory activity may be related to the activation of AMPK pathway.