4-Methylcoumarin Derivatives with Anti-inflammatory Effects in Activated Microglial Cells

Inflammation contributes to the pathogenesis of neurodegenerative diseases and anti-inflammatory compounds may have a role in prevention or treatment of these pathologies. 4-Methylcoumarins are effective antioxidants with anti-inflammatory properties. In this study, the inhibitory effects of two 4-methylcoumarin derivatives, 7,8-dihydroxy-3-ethoxycarbonylmethyl-4-methylcoumarin (DHEMC) and 7,8-diacetoxy-3-ethoxycarbonylmethyl-4-methylcoumarin (DAEMC) were examined on the inflammatory processes induced by lipopolysaccharide (LPS) in activated primary rat microglial cultures. LPS-induced production of nitric oxide (NO, measured by Griess method) and other pro-inflammatory mediators, thromboxane (TX) B-2 and prostaglandin (PG) E-2 (both determined by radioimmunoassay (RIA)), as well as tumor necrosis factor (TNF)-alpha (determined by enzyme-linked immunosorbent assay (ELISA)) were inhibited in the presence of 100 mu m DHEMC and DAEMC. DAEMC was able to significantly inhibit NO, TXB2 and TNF-alpha production also at 50 mu M. Both compounds at 100 mu M significantly lowered cyelooxygenase-2 (COX-2) protein expression in LPS-stimulated microglial cells measured by Western blot, but only DAEMC showed an inhibitory effect on inducible nitric oxide synthase (iNOS) protein expression at 100 mu M. In conclusion, our findings show that 4-methylcoumarin derivatives can modulate inflammatory pathways in microglial cells, probably by acting at the protein expression level.