Effects Of Short-Term High Sugar Diet On Th1/Th2 And Th17/Treg Balance In Asthmatic Mice

Objective By giving asthma mice short-term high sugar diet to detect the changes of airway inflammation and the expression levels of related inflammatory cells,cytokines and proteins,and to explore the effects of short-term high sugar diet on airway inflammation and Th1/Th2 and Th17 /Treg balance.MethodBALB/C female mice at 6-8 weeks were randomly divided into four grou ps: control(NORMAL)group,high sugar diet(HSD)group,asthma(Ovalbumi n,OVA)group,and high sugar diet/asthma(HSD/OVA)group.The NORMAL group were given ordinary drinking water,the HSD group were given 20% glu cose drinking water,The model of asthma was established by ovalbumin and were given ordinary drinking water in OVA group,the HSD/OVA group were gi ven 20% glucose drinking water,and the asthma model was established in the same way,the behavior and temperament changes of the mice were observed during feeding,and all mice were killed on the 42 nd day.Hematoxylin and eosi n(HE)and periodic acid-schiff(PAS)were used to observe the changes of lun g inflammation and goblet epithelial cell proliferation.The total cells of alveol ar lavage fluid were counted,and reichsen staining was used to count eosinoph ilic granulocyte percentage.The expression level of Immunoglobulin E(IgE)in serum of mice was detected by Enzyme linked immunosorbent assay(ELISA).Flow cytometry was used to analyze the levels of Th1、Th2、Th17、Treg cel ls in mouse spleen.The Polymerase Chain Reaction(PCR)was used to detect the expressions of IFN-γ,IL-4,IL-17,TGF-β and IL-10 cytokines in lung tiss ue.The Immunohistochemistry(IHC)was to assess protein expression levels of IL-4,IL-17,IL-10,IFN-γ,TGF-β and FOXP3 in lung tissue.Result1.Compared with NORMAL group,HSD group mice show no obvious symptoms of acute asthma attack and no obvious inflammatory cells infiltration or goblet cell proliferation in lung tissue in HE and PAS staining result,there is no significant changes in the total number of cell and eosinophil count in alveolar lavage fluid(P > 0.05).There is no significant difference in serum specific IgE level of mice(P > 0.05).In the OVA and HSD/OVA group,The mice are nasal scratching repeatedly and irritability occurred in the process of atomization,and inflammatory infiltration around the lung tissue are obvious,goblet cells proliferation were visible.The total number of cell and percentage of eosinophilic granulocyte in alveolar lavage fluid are significantly increased(P < 0.05),and the specific IgE in ocular blood of mice is significantly increased(P < 0.05).Compared with the OVA group,the inflammatory infiltration of the lung in HSD/OVA group are more obvious,a large number of goblet cells formation and mucus secretion could were be seen,the number of eosinophils is increased more significantly(P < 0.05),the total number of alveolar lavage fluid and the specific IgE of the mice is not increased significantly(P > 0.05).2.Compared with NORMAL group,the proportions of Th1,Th2,Th17 and Treg cells in spleen of mice in HSD group are not significantly changed(P > 0.05);The proportion of Th2 and Th17 cells in OVA and HSD/OVA groups are significantly increased(P < 0.05),and the proportion of Th1 cell and Treg cells are significantly decreased(P < 0.05).The increase of Th2 and Th17 cels in HSD/OVA group are more significant than that in OVA group(P < 0.05),and the proportion of Th1 cell is decreased more significantly than that in OVA group(P < 0.05),the proportion of Treg cells is decreased,the difference is not significant(P > 0.05).3.Compared with NORMAL group,mRNA relative expression levels of IL-4、IL-17、IL-10 and TGF-F-β in lung of mice in HSD group have no significant change(P > 0.05),IFN-γ is decreased significantly(P< 0.05).The mRNA relative expressions of IL-4 and IL-17 are significantly increased in OVA and HSD/OVA group(P < 0.05),the mRNA relative expressions of TGF-β and IFN-γ are significantly decreased(P <0.05).Compared with the OVA group,the mRNA relative expression levels of IL-4、IL-17 and IL-10 in HSD/OVA group are significantly increased(P < 0.05),the mRNA relative expression level of TGF-β is significantly decreased(P < 0.05),the mRNA relative expression level of IFN-γ is not significantly decreased(P > 0.05).4.Compared with NORMAL group,the protein levels of IL-4、IL-17、IL-10、IFN-γ、TGF-β and FOXP3 in lung of mice in HSD group have no significant change;The expression levels of IL-4 and IL-17 in OVA and HSD/OVA group are significantly increased(P < 0.05),the protein levels of IFN-γ,TGF-β and FOXP3 are significantly decreased(P < 0.05),the level of IL-10 is not significantly change(P >0.05).The levels of IL-4 and IL-17 in HSD/OVA group are significantly increased compared with those in OVA group(P < 0.05),while the level of TGF-β is significantly decreased(P < 0.05),the levels of IL-10、IFN-γ and FOXP3 are not significantly change(P > 0.05).Conclusion The proportion of Th1/Th2 and Th17/Treg cells are unbalanced in asthmatic mice.Short-term high-sugar diet can regulating the expression of cytokine such as IFN-γ,IL-4,IL-17,TGF-β,IL-10 and FOXP3 protein to affect the balance of Th1/Th2 and Th17/Treg cells in asthmatic mice,to aggravated airway inflammation in asthmatic mice.