Blockade of nuclear factor-κB signaling pathway and anti-inflammatory activity of cardamomin, a chalcone analog from Alpinia conchigera

Nuclear factor-kappa B (NF-kappa B) and the signaling pathways that regulate its activity have become a focal point for intense drug discovery and development efforts. NF-kappa B regulates the transcription of a large number of genes, particularly those involved in immune, inflammatory, and antiapoptotic responses. In our search for NF-kappa B inhibitors from natural resources, we identified cardamomin, 2', 4'-dihydroxy-6'-methoxychalcone, as an inhibitor of NF-kappa B activation from Alpinia conchigera Griff (Zingiberaceae). In present study, we demonstrated the effect of cardamomin on NF-kappa B activation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and LPS-induced mortality. This compound significantly inhibited the induced expression of NF-kappa B reporter gene by LPS or tumor necrosis factor (TNF)-alpha in a dose-dependent manner. LPS-induced production of TNF-alpha and NO as well as expression of inducible nitric-oxide synthase and cyclooxygenase-2 was significantly suppressed by the treatment of cardamomin in RAW264.7 cells. Also, cardamomin inhibited not only LPS-induced degradation and phosphorylation of inhibitor kappa B alpha (I kappa B alpha) but also activation of inhibitor kappa B (I kappa B) kinases and nuclear translocation of NF-kappa B. Further analyses revealed that cardamomin did not directly inhibit I kappa B kinases, but it significantly suppressed LPS-induced activation of Akt. Moreover, cardamomin suppressed transcriptional activity and phosphorylation of Ser536 of RelA/p65 subunit of NF-kappa B. However, this compound did not inhibit LPS- induced activation of extracellular signal-regulated kinase and stress-activated protein kinase/c-Jun NH2-terminal kinase, but significantly impaired activation of p38 mitogen-activated protein kinase. We also demonstrated that pretreatment of cardamomin rescued C57BL/6 mice from LPS-induced mortality in conjunction with decreased serum level of TNF-alpha. Together, cardamomin could be valuable candidate for the intervention of NF-kappa B-dependent pathological condition such as inflammation.