Objectives: Rehmannia glutinosa pharmacopuncture solution (RGPS) was investigated to determine both its anti-allergic inflammatory effects on mast cells and its detailed mechanism of actions.
Methods: We investigated whether RGPS suppress cytokines, enzymes, FcεRI expression and FcεRImediated signaling in RBL-2H3 cells stimulated with anti-DNP IgE/DNP-HSA. The suppressive effects of RGPS on the levels of cytokines such as IL-1β, IL-6 and GM-CSF were measured using emzyme-linked immunospecific assay (ELISA). The mRNA expression levels of cytokines, enzymes (HDC2, COX-1, COX-2 and 5LO) and FcεRI αβγsubunits were measured using reverse transcription polymerase chain reaction (RTPCR) method. The activation of FcεRI-mediated signaling was examined using Western blot analyses.
Results: RGPS suppressed production of proinflamm-atory cytokines (IL-1β, IL-6, and GM-CSF) in stimulated RBL-2H3 cells significantly (p< 0.05). RGPS also suppressed mRNA expression of inflammatory enzymes (HDC2, COX-1, COX-2, 5LO). In addition, mRNA expression levels of FcεRIα, FcεRIβand FcεRIγ were lowered by treatment with RGPS. Finally, RGPS prevented phosphrylation of Lyn, Syk, LAT, Gab2, PLC γ1/2, PI3K, Akt, cPLA2 and IκBα.
Conclusions: RGPS effectively suppresses mast cell activations such as degranulation and inflammatory response via down-regulation of the FcεRI-mediated signaling pathways in IgE/Ag-stimulated mast cells.
Keywords: FcεRI expression; FcεRI mediated signaling; Rehmannia glutinosa pharmacopuncture solution (RGPS); cytokine; enzyme.