科研之友
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摘要
Fc汍RI, The high affinity receptor for IgE plays a critical role in triggering the allergic reactions. It is responsible for inducing mast cell degranulation and deliberation of allergy mediators when it is aggregated by allergen and IgE complexes. Fc汍RI on the mast cells consists of three subunits; 汐 chain directly binds IgE, 汕 chain and dimmer of 污 chains together mediate intracellular signaling. Cross-linking of IgE-bound Fc汍RI on the surface of mast cells and basophils by the multivalent antigen induces release of chemical mediators. The present in vitro study was designed to investigate the effect of synthetic Fc汍RI-汐 siRNA on the antigen-induced activation of MC/9 cells. MC/9 cells which are murine mast cells were transfected by Fc汍RI-汐 siRNA and negative control siRNA. After 6 h, anti-DNP (Dinitrophenyl) IgE was used for the cells sensitization. Then the cells were challenged with Dinitrophenyl-Human Serum Albumin (DNP-HSA) for mast cell degranulation induction before collection of supernatants. The amount of mRNA and protein expression was measured by Real Time PCR and western blot analysis, respectively. Determination of the expression rate of Fc汍RI-汐 on cell surface was achieved by flow cytometry. ELISA and spectrophotometry methods were used subsequently for measuring the effects of Fc汍RI-汐 siRNA on antigen-induced histamine and 汕-hexosaminidase release. Fc汍RI-汐 siRNA treated cells showed significant decrease in Fc汍RI-汐 mRNA and protein expression in comparison to control cells. Fc汍RI-mediated mast cell release of 汕-hexosaminidase and histamine were also inhibited. In this study it was shown that Fc汍RI-汐 siRNA could suppress Fc汍RI-汐 expression and inhibited degranulation and histamine release in antigen-stimulated MC/9 cells. In conclusion, knock-down of Fc汍RI-汐 by siRNA could be a promising method for inhibition of the mast cell-mediated allergic reactions.
