Objective: Modified-Okbyungpoongsan(KOB03) is a polyherbal medicine derived from an Korean Medicine prescription, Okbyungpoongsan which is traditionally used to treat for the patients with respiratory diseases such as asthma, allergic rhinitis, and cold. In the present study, we investigated the effects of KOB03 on allergic asthma, and the action mechanism in mast cell activation.
Materials and Methods: For in vitro study, RBL-2H3 mouse basophilic mast cells were cultured and treated with KOB03 at different concentration for indicated time. After the stimulation with compound 48/80 (C48/80) or DNP-IgE/HSA (IgE/Ag) in the cells, the degranulation was observed by microscope after toluidine blue staining, and the levels of histamine and β-hesoximinidase were measured in culture supernatant by EIA. The expression of inflammatory cytokines, TNF-α, IL-4, IL-13 and IFN-γ was detected by RT-PCR, and the phosphorylation of ERK, JNK, and p38 MAPK was determined by Western blot, respectively. For in vivo study, allergic asthma animal model was prepared using ovalbumin (OVA) sensitized/challenge in mice. KOB03 was administrated orally in allergic asthma mice during OVA challenge for 7 days once a day. The levels of OVA-specific IgE, and allergic inflammatory cytokines (TNF-α, IL-4, IL-5, and IL-13) were measured in bronchoalveolar lavage fluids (BALF) and sera of mice by ELISA. The infiltrated numbers of inflammatory cells in BALF were also counted after stain with toluidine blue and diff-quik dye. The histological changes of lung tissues were observed by hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) and Mansson’s trichrome (M-T) staining.
Results: In RBL-2H3 cells, the treatment of KOB03 at 0.5 mg/mL was significantly inhibited IgE/Ag induced release of β-hexosaminidase and histamine through inhibition of the cell degranulation. KOB03 at 0.5 mg/mL also significantly inhibited the expression of TNF-α, IL-4 and IL-13 mRNA in IgE/Ag-stimulated RBL-2H3 cells. KOB03 at 0.1 mg/mL and 0.5 mg/mL significantly suppressed the phosphorylation of ERK1/2 MAPK in IgE/Ag-stimulated cells. In allergic asthma mice, the administration of KOB03 at 200 mg/kg significantly decreased the release of OVA-specific IgE, TNF-α, IL-4, IL-5, and IL-13 in serum and BALF of mice. The KOB03 administration was significantly decreased the infiltration of inflammatory cells including total cells, T cells, macrophages, mast cells, and eosinophils, and inhibited the mucin accumulation and fibrosis in bronchiole and lung tissues of allergic asthma mice.
Conclusion: These results indicate that KOB03 has a therapeutic potential on allergic asthma through regulation of the Th2-mediated allergic response and inflammation, and its mechanism is underly the regulation of mast cell activation.