Andrographolide interferes with binding of nuclear factor-κB to DNA in HL-60-derived neutrophilic cells

1 Andrographolide, the major active component from Andrographis paniculata, has shown to possess anti-inflammatory activity. Andrographolide inhibits the expression of several proinflammatory proteins that exhibit a nuclear factor kappa B (NF-kappa B) binding site in their gene. 2 In the present study, we analyzed the effect of andrographolide on the activation of NF-kappa B induced by platelet-activating factor (PAF) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) in HL-60 cells differentiated to neutrophils. 3 PAF (100 nM) and fMLP (100 nM) induced activation of NF-kappa B as determined by degradation of inhibitory factor B alpha (I kappa B alpha) using Western blotting in cytosolic extracts and by binding to DNA using electrophoretic mobility shift assay (EMSA) in nuclear extracts. 4 Andrographolide (5 and 50 mu M) inhibited the NF-kappa B-luciferase activity induced by PAF. However, andrographolide did not reduce phosphorylation of p38 MAPK or ERK1/2 and did not change IkBa degradation induced by PAF and fMLP. 5 Andrographolide reduced the DNA binding of NF-kappa B in whole cells and in nuclear extracts induced by PAF and fMLP. 6 Andrographolide reduced cyclooxygenase-2 (COX-2) expression induced by PAF and fMLP in HL-60/neutrophils. 7 It is concluded that andrographolide exerts its anti-inflammatory effects by inhibiting NF-kappa B binding to DNA, and thus reducing the expression of proinflammatory proteins, such as COX-2.