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  • Title: Regulation of inflammatory mediators in lipopolysaccharide-stimulated RAW 264.7 cells by 2''-hydroxy-3''-en-anhydroicaritin involves down-regulation of NF-kappaB and MAPK expression.
    Author: Ci X, Liang X, Luo G, Yu Q, Li H, Wang D, Li R, Deng X.
    Journal: Int Immunopharmacol; 2010 Sep; 10(9):995-1002. PubMed ID: 20462523.
    Abstract:
    2''-hydroxy-3''-en-anhydroicaritin, a flavone, was isolated from the Chinese medicinal herb Epimedium brevicornum for the first time. In our previous study, we have carried out a screening program to identify the anti-inflammatory potentials of 2''-hydroxy-3''-en-anhydroicaritin. In the present study, we further found that this compound regulated lipopolysaccharide (LPS)-induced levels of nitric oxide (NO), and prostaglandin E(2) (PGE(2)) (**p<0.01 or *p<0.05), and reduced levels of iNOS and COX-2 in RAW 264.7 macrophages in a concentration-dependent manner. We further investigated signal transduction mechanisms to determine how 2''-hydroxy-3''-en-anhydroicaritin affects RAW264.7 macrophages pretreated with 0.5, 2.5, or 12.5mg/L of 2''-hydroxy-3''-en-anhydroicaritin 1h prior to treatment with 1mg/L of LPS. Thirty minutes later, cells were harvested and mitogen-activated protein kinases (MAPK) activation and I kappaB alpha were measured by western blotting. Alternatively, the macrophages were fixed and nuclear factor-kappaB (NF-kappaB) activation was measured by immunocytochemical analysis. Signal transduction studies showed that the flavone significantly inhibited extracellular signal-regulated kinase (ERK), p38, and c-jun NH2-terminal kinase (JNK) phosphorylation protein expression. The flavone also inhibited p65-NF-kappaB translocation into the nucleus by I kappaB alpha degradation. Therefore, 2''-hydroxy-3''-en-anhydroicaritin may inhibit LPS-induced production of inflammatory cytokines by blocking NF-kappaB and MAPK signaling in RAW264.7 cells.
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