| Objective:The study was carried out by establishing the models of allergic rhinitis group and control group with the mice which were knocked out mi RNA-155(mi R-155)gene and BALB/c WT mice.Then discuss the effects of mi RNA-155 how to regulate and control the trend of GATA3,Th1/Th2 and type 2 innate lymphocyte(ILC2).We hope that these results will provide a further explanation of complex pathogenesis and some theoretical basis for the diagnosis and treatment of AR in the future.Methods:(1)Purchased mi RNA-155 KO mice and BALB/c WT mice,respectively,and used the OVA and PBS to established AR model and control group model of mice,and there were eight mice in each group;(2)Weighing each mouse after the last challenge and observe the behavioral change of each mouse such as scratching nose,sneezing and runny nose for 30minutes;(3)Taking the nasal tissues of mice and make pathological sections,and make HE staining and PAS staining to observe the number of eosinophils cells and goblet cells in the nasal mucosa of each group,in addition,the arrangement of cilia in the nasal mucosa epithelium;(4)Collecting the venous blood of mice through the inner canthus,then extract the serum,test the concentration of IL-13,IL-33,OVA s Ig E and IFN-γ by ELISA;(5)Peeling off the nasal mucosa of mice,the extract RNA,proceed reverse transcription and amplification,then compare the relative expressions level of mi RNA-155,GATA3,IL13 and IFN-γ in nasal mucosa of each group lastly;(6)Preparing the single cell suspension of spleen,then detect and compare the Th1/Th2 and the ratio of ILC2 by flow cytometry.Results:(1)After observing the last challenge of nasal drip,weighing mice and observing the behavioral,the weight difference between groups of mice had no significant effect on the research indexes(P>0.05),then statistics the times of scratch nose,sneezing and the condition of runny nose,calculating the behavioral scores of each mice,the score of AR group was higher than mi RNA-155 KO group and control group,and there is a statistical difference(P < 0.05);(2)When the pathological section was stained with HE,we count the number of eosinophils in the AR group was more than mi RNA-155 KO mice and control group,and there is a statistical difference(P<0.05),and the cilia of AR mice were more disordered and more interrupted.And while the pathological section was stained with PAS,we found that in AR mice the number of goblet cell is more than other groups,and there is a statistical difference(P<0.05);(3)ELISA results showed that the serum concentrations level of IL-13,IL-33 and OVA s Ig E of AR group were higher than mi RNA-155 KO group and control group,while the IFN-γ serum concentration of AR group was lower than control group,and slightly higher than mi RNA-155 KO group;(4)The expression level of mi RNA-155,GATA3-3 and IL-13 in nasal mucosa of mice in AR group was higher than that in the mi RNA-155 KO group and control group,while the expression level of IFN-γ in AR group was lower than that in other two groups;(5)The flow cytometry test the single cell of spleen,results showed that the proportion of Th2 cells and ILC2 cells in AR group was higher than that in mi RNA-155 KO group and control group,while the proportion of Th1 was lower than that in control group,and higher than that in KO group,in short,the Th1/Th2 ratio in AR group was lower than that in control group and KO group.Conclusion:(1)In the research of AR,mi RNA-155 may promote the predominance differentiation of CD4+T cells to Th2,reduce the Th1/Th2 ratio,and enhance the differentiation of ILC2 at the same time.Besides,mi RNA-155 also promote the generation of Th2 cytokines and the development of AR;(2)Knockout the mi RNA-155 of mouse can inhibit the expression of GATA3 and differentiation of Th2 and ILC2,then relieve the symptoms of AR. |
PDF Full Text Request