Effects Of Recombinant UBC13 Protein On Th2,Th17 Polarization And Related Pathways In Mice Bronchial Asthma Model

Objective: The purpose of the experiment was to construct a suitable asthma model to explore the effects of recombinant UBC13 protein on Th2,Th17 polarization and related pathways.Methods:（1）Selection of bronchial asthma model: 24 BALB/c mice were randomly divided into PBS group,LPS modeling group,and Al（OH）₃ modeling group.The two modeling groups were injected with the corresponding sensitizing solution on day 0,7,and 14 respectively.On day 21,The two modeling groups were stimulated with 5 %OVA solution for for 7 days,30 minutes each.The PBS group was sensitized and nebulized with PBS instead of the sensitizer at the same time point.Both of changes in serum IgE content,lung pathological sections and the effects of CD4⁺ T lymphocytes of each group were determine to whether the modeling was successful.（2）Effect of recombinant UBC13 protein on bronchial asthma models: According to the first part of the LPS modeling method,24 BALB/c mice were randomly divided into PBS group,asthma model group,UBC13 control group,and positive control group（budesonide）.Record the weight changes of mice in each group,the serum IgE content of the mice by ELISA.HE and PAS staining observed the histological sections of lungs.Real-time quantitative PCR and ELISA detected the expression of Th2 and Th17 cytokines.Immunohistochemistry detected the expression of transcription factors GATA3 and RORγt.Western Blot detected the expression of key proteins IκBα,p-IκBα,and p65 in the NF-κB signaling pathway.Results:（1）Histopathological observation showed that airway inflammation aggregated,airway smooth muscles became thicker,and mucus secretion occurred in both models.Compared with the PBS group,the serum IgE content of both models was significantly increased（P<0.001）,the expression of CD4⁺ T lymphocytes was significantly increased（P <0.05）.（2）ELISA detection of serum IgE content showed that the IgE content of the model group and the UBC13 protein group was significantly increased（P<0.01）.Compared with the model group,the IgE content of the protein group and the positive control group were significantly decreased（P<0.01）.Lung pathological section results showed that airway inflammation gathered and a large amount of mucus secretion.The inflammation and mucus secretion in the UBC13 group and the positive group were significantly improved.The q PCR results showed that compared with the PBS group,the Th2 cytokines（IL-4,IL-5,and IL-13）m RNA expression in the lung tissue of the model group were significantly increased（P<0.01）.Th17 cytokines（IL-1β,IL-6,IL-17 A,and IL-23）m RNA expression levels were extremely significantly increased（P<0.001）,compared with the model group,while Th17 cytokines m RNA expression in the UBC13 group and the positive control group were significantly reduced（P<0.05）.In alveolar lavage fluid（BALF）,ELISA results showed that compared with the PBS group,IL-4 and IL-5 secrection in model group were significantly increased（P<0.001）,IL-17 A secrection was significantly increased（P<0.01）;compared with the model group,IL-4 and IL-5 secrection in the UBC13 group and the positive group were significantly reduced（P<0.001）,and IL-17 A secrection was not significantly difference（P>0.05）.Immuno-histochemical results showed that compared with the PBS group,the expression levels of GATA3 and RORγt in the model group were significantly increased（P<0.01）,compared with the model group,the UBC13 group and the positive group were significantly reduced（P<0.05）.Western Blot results showed that compared with the PBS group,the expression levels of p-IκBα and nucleus p65 in the model group were significantly increased（P<0.01）,compared with the model group,the expression levels of cytoplasmic p-IκBα and nucleus p65 in the UBC13 group and the positive group were significantly reduced（P<0.01）,and the expression levels of cytoplasmic IκBα and p65 were significantly increased（P<0.01）.Conclusion: The asthma model constructed with LPS as an adjuvant is more severe,and the phenomenon of neutral mucus secretion is more in line with the needs of the next test.UBC13 protein can reduce the accumulation of airway inflammation in asthma models by inhibiting the activation of NF-κB signaling pathway,suppress the polarization of Th2 and Th17 cells,and perform certain therapeutic effects on airway inflammation in asthma models.