Anti-inflammatory Activity of Hangeshashinto in IL-1β-stimulated Gingival and Periodontal Ligament Fibroblasts

Although a large number of Kampo, Japanese traditional medicines, have been used for the treatment of oral diseases, little is known on their relative potency and endotoxin contamination. In order to obtain basic data for clinical applications, 10 Kampo, and 25 constituent plant extracts were tested for the contamination of lipopolysaccharide (LPS)-like substances, and anti-inflammatory activity. Human gingival (HGF) and periodontal ligament fibroblasts (HPLF) were cultured in 10% fetal bovine serum supplemented with Dulbecco's modified Eagle's medium. Viable cell number was measured by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Prostaglandin (PGE(2)) was determined by enzyme immunoassay. Cyclo-oxygenase (COX)-1 and COX-2 protein expressions were determined by western blot. COX activity was measured using Cox Inhibitor Screening Assay Kit. LPS, quantified by Endotoxin assay kit, was undetectable or relatively low in the test samples except for rikkosan and unseiin. Hangeshashinto potently inhibited PGE(2) production by interleukin (IL)-1 beta-stimulated HPLFs and HGFs. Hangeshashinto suppressed the expression of COX-2 protein, but not that of COX-1 protein in IL-1 beta-induced HGF cells. Hangeshashinto slightly, but not significantly, inhibited both COX-1 and COX-2 activity. The present study provides the basis for clinical application of hangeshashinto for the treatment of stomatitis.