Inhibitory effect of extracts of hainosan ( painongsan ) and its constituent crude drugs on production of monocyte chemoattractant protein‐1 and interleukin‐6 in murine fibroblasts treated with Porphyromonas gingivalis

Aim: Hainosan (painongsan), composed of Platycodon grandiflorum root (PG), Paeonia lactiflora root (PL), and Citrus aurantium immature fruit (AF), is a formula used in traditional Japanese (Kampo) and Chinese medicine to treat purulent diseases, including gingivitis. In this study, we investigated the anti-inflammatory effects of hainosan extract (HNS) using both traditional direct and serum pharmacological techniques.Methods: BALB/c mice were orally treated with HNS or extracts of its constituent crude drugs for one week. Murine RCB2767 fibroblasts were incubated with a medium containing heat-killed Porphyromonas gingivalis and HNS, extracts of the constituent crude drugs, or serum samples collected from mice orally treated with these extracts. The monocyte chemoattractant protein 1 (MCP-1) and interleukin (IL)-6 in the medium were measured by enzyme-linked immunosorbent assay (ELISA). The effect of HNS and its constituent crude drugs on the proliferative potential of fibroblasts was evaluated by radioisotope-labeled thymidine uptake capacity.Results: The concentrations of MCP-1 and IL-6 in the culture supernatants were significantly increased by treatment with heat-killed bacteria. In addition, PG and PL extracts significantly inhibited MCP-1 and IL-6 production. Furthermore, supplementation of serum samples from mice orally treated with the extracts to the culture medium resulted in significant inhibition of induction of these cytokines. HNS, extracts of PG, PL and AF, and serum samples collected from mice treated with these extracts significantly augmented H-3-thymidine uptake in fibroblasts.Conclusion: Our results demonstrated that HNS has anti-inflammatory effects in murine fibroblasts and that PL and PG could contribute to the effects of hainosan.