Effects Of Neohesperidin In Degranulation Of RBL-2H3 Cells And Allergic Asthma Disease Induced By OVA

Food allergy is one of the most common allergic diseases which associated with nausea,vomiting,diarrhoea,allergic asthma,allergic dermatitis,allergic rhinitis and allergic shock.Its incidence manifests a rising tendency with each passing year,caused a heavy burden for patient's family and the society.Neohesperidin is a kind of dihydroflavone glycoside,which belongs to the metabolites of citrus plants in rutaceae.Neohesperidin is widely distributed in various plants.Neohesperidin exhibit various biological activities,such as anti-oxidant,anti-inflammatory,anti-tumor,anti-bacterial and the protection of the cardiovascular system.However,the anti-asthma activitity of neohesperidin induced by food allergy has yet to be reported.In the present study,we investigated the possible anti-allergic activities of neohesperidin in vivo and in vitro as well.To do this,the effect of neohesperidin on degranulation of RBL-2H3 cells induced by antigen and allergic asthma mice induced by OVA were examined.The specific research content is as follows.1.In vitro experiments:The viability of RBL-2H3 cells was determined using an MTT assay.The release of?-hexosaminidase from antigen-stimulated RBL-2H3 cells was measured.The effect of neohesperidin on antigen-induced intracellular Ca2~+mobilization in RBL-2H3 cells was measured with fluorescence microscope.The results show that the survival rate of RBL-2H3 cells was maintained above 90%treated with different concentrations of neohesperidin and the viability of RBL-2H3 cells did not decrease with the increasing of neohesperidin concentration,indicating that neohesperidin has no significant effect on RBL-2H3 cells viability.Different concentrations of neohesperidin has a inhibitory effect on the release of?-hexosaminidase in antigen-stimulated RBL-2H3 cells,and the inhibitory effect shows the best in the concentration of 3.13?mol/L.The neohesperidin show a suppressive effect on antigen-induced intracellular Ca2~+mobilization in RBL-2H3 cells.2.In vivo experiments:Allergic asthma model mice was established and then obtain the samples.The weight as well as the index of thymus and spleen was recorded and calculated.The levels of IgE and IgG1 in serum were measured by ELISA.The production of Th2cytokines(IL-4 and IL-5)and Th1 cytokines(IFN-?)in BALF were also quantified by ELISA.The percent of CD4~+and CD8~+T cells in the spleen were detected by flow cytometry method.The infiltration of inflammatory cells in peribronchial areas,inflammatory cells in lung tissues were conducted with the hematoxylin and eosin staining(H&E).The results show that there was no obvious change of neohesperidin in the levels of IgE and IgG1 in serum.However,neohesperidin reduced the levels of IL-4 as well as IL-5 and increased the level of IFN-?in BALF of mice.Neohesperidin reduced the number of CD4~+T cells and increased the number of CD8~+T cells in the spleen of mice.Neohesperidin also reduced the infiltration of eosinophils in lung tissue,changed the infiltration of inflammatory cells in the bronchioles,peribronchial areas and intraluminal.