| Objective:To investigate the effects of erythropoietin(EPO)on regulatory T cells(Treg)and helper T cell 17(Th17)-related cytokines IL-10、TGF-β、IL-17 and pathological changes in the lungs which uses the ovalbumin-induced to acute exacerbation asthma mouse model.Further explored the role of EPO in the asthmatic mouse model and its intervention mechanism,providing a useful experimental basis for the clinical application of EPO.Methods:To chosse a total of 45 male clean grade BALB/c mice aged 6-8 weeks and weighing approximately 18-22 g were randomly divided into five groups:normal control group,asthma model group,dexamethasone group,low-dose EPO group and high-dose EPO group.Then were sequentially labeled with capital letters A through E,with 9 mice in each group.The first step of the experiment was sensitization,beginning on the first day and then sensitized once every 7 days and then sensitized 3 times.During the sensitization process,four groups of B,C,D,and E were treated with 200 ul phosphate buffer solution(Phosphate Buffered Saline,PBS)(10 ug ovalbumin OVA+2 mg aluminum hydroxide Al(OH)3 adjuvant).Intraperitoneal injection.The second step of antigen challenge was calculated with the last sensitization,followed by 2.5%OVA inhalation on the 8th day,continuous nebulization for 7 days,and each nebulization lasting 1/2 hour.In the third step,dexamethasone(1 mg/kg)was injected back into the C group 1/2 hour prior to each atomization.In groups D and E,the low concentration of EPO(1000 IU/kg)and high concentrations of EPO were injected in the same manner.2000IU/kg).After the challenge,the mice showed symptoms of asthma,such as scratching their ears,wheezing,restlessness and restlessness,indicating that the asthma model was successfully prepared.In this experiment,group A was replaced with normal saline,and the injection method and time were the same.Enzyme-linked immunosorbent assay(ELISA)was used to determine the interleukin-10(IL-10),erum transforming growth factor-β(TGF-β)in regulatory T cells(Treg)and Levels of IL-17,an important cytokine of helper T cells 17(Th17),Bronchoalveolar lavage fluid counted by a Wright-Giemsa microscope The number of airway inflammation-associated eosinophile granulocyte(EOS)was measured.The pathological changes of lung tissue were observed under a microscope after HE staining.Results:(1)Levels of IL-10 and TGF-β:Compared with the levels of IL-10 and TGF-P in group A,the levels of IL-10 and TGF-β in group B decreased,and the difference was statistically significant(P<0.05);Compared with the levels of IL-10 and TGF-β in group B,the levels of C,D,and E in the three groups were all increased,the difference was statistically significant(P<0.05);different concentrations of EPO Comparison between groups:D group was higher than E group,the difference was not statistically significant(P>0.05);(2)IL-17 level and EOS count:compared with A group IL-17 level and EOS count The levels of IL-17 and EOS counts in group B were significantly higher(P<0.05);the IL-17 levels and EOS counts in group B were compared with those in groups C,D,and E.The level of concentration and count decreased,and the difference was statistically significant(P<0.05).The differences between different EPO concentration groups were compared:D group was lower than E group,the difference was not statistically significant(P<0.05);(3)HE staining After the microscope,the changes of the lung tissue of the mice were compared:there was no pathological change in the lung tissue of the mice in the group A;the histomorphology of the lung tissues in the mice of the group B changed significantly.Pulmonary alveolar rupture fusion,its interval hypertrophy,diffuse inflammatory cells can be seen in all layers of the wall,mucosal epithelial loss,cilia collapse or even disappear,basement membrane thickening;C,D,E three groups of mouse lung tissue under the microscope Morphological changes were observed.Compared with group B,the changes of microscopic structure were lighter than those of group B.The histopathological changes of lung tissue in groups C,D,and E were lighter than those in group C.Compared with the E group,the difference is not obvious.Conclusion:Changes in serum levels of IL-10,TGF-β,and IL-17 in a mouse model of bronchial asthma suggest a Treg/Thl7 imbalance.Administration of appropriate concentrations of exogenous EPO may have an inhibitory effect on acute exacerbation of bronchial asthma.The mechanism may be through the regulation of serum IL-10,TGF-β,and IL-17 levels to restore the balanced Treg/Th17 ratio and reduced eosinophilic airway inflammation to achieve. |
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