Anti-inflammatory effect of Sinhyowoldo-san Extract with regard to Pro-inflammatory Mediators in PMA plus A23187-induced Human Mast Cells

Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea.But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood.To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1).Methods : In the present research, cell viability was measured by MTS assay.pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-к B).The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and NF-к B in PMA+A23187-activated HMC-1 cells.Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and 250 µg/ml).SHWDS (250 µg/ml) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells.Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression.In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2).Then, SHWDS suppressed activation of nuclear factor NF-к B in nuclear, degradation of IkB α in cytoplasm.Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and NF-к B activation, thereby decreasing the expression of pro-inflammatory genes.