| Objective:Through the detection of the number of CD4+CD25+regulatory T cells (CD4+of CD25+of Treg) or Th17cells and t theexpression level of IL-10and IL-17,in asthma and after glucocorticoid orgrass mycobacterial treatment, analyze and explore the mechanism of CD4+CD25+regulatory T cells and Th17cells in the pathogenesis of asthma;explore the impact of CD4+CD25+regulatory T cells/Th17cells immunebalance after the Mycobacterium phlei treatment.Methods:Specificpathogen-free female BALB/c mice were24,7weeks old and weighingabout23-25g, the random number method table completely randomly dividedinto four groups, normal control group, asthma group, dexamethasonetreatment asthma group, Mycobacterium phlei treatment of asthma group. Ondays1and14days of experimental asthma group, asthma, dexamethasonetreatment group and of asthma Mycobacterium phlei treatment group mice byintraperitoneal injection of0.2ml of ovalbumin (OVA) and aluminumhydroxide (Al (OH)3) mixture [containing OVA100ug, AL (OH)32mg]allergens. In the first21-25days of the experiment, these three groups ofOVA-sensitized mice were placed in a homemade spray box to box, tostimulate OVA concentration of2%liquid atomization, atomization30minutes a day. Asthma dexamethasone stimulate30minutes before the intraperitoneal injection of dexamethasone treatment group each atomization,injection volume1mg/Kg continuous infusion for five days. AsthmaMycobacterium phlei treatment group in the experiment20days23days26days by intraperitoneal injection Mycobacterium phlei, a total of threeinjections, each injection are0.57ug. In the first27-55days of the experimentcontinue to be2%OVA to stimulate liquid atomization, and atomization each30minutes three times a week. Normal control group were givenintraperitoneal saline and saline spray. All mice in the last challenge,24h eyeblood, hoping the mice were sacrificed, the line of mouse alveolar lavage,Bronchoalveolar lavage fluid spare. Mice left lung for biopsy was observed inmice lung airways and the inflammatory changes. Mice spleen preparation ofmouse spleen mononuclear cell suspension. The respective percentages offlow cytometry each group of mice spleen mononuclear cell suspensions ofCD4+T cell subsets in the CD4+CD25+regulatory T cells and Th17cells.ELISA for detection of cytokines in the serum and bronchoalveolar lavagefluid IL-10, IL-17expression levels.Results:1) mice with ovalbumin aerosolto stimulate less dynamic irritability, shortness of breath or difficultybreathing, lips cyanotic, unresponsive, the performance of incontinence, suchas acute asthma attacks; Pathological examination showed: the bronchiolesand blood vessels around showed significantly eosinophils, lymphocytesincreased inflammatory cell infiltration consisting mainly of bronchial wallthickening, luminal narrowing, some of the bronchiolar epithelial cell shedding incomplete, suggesting that the mouse asthma model wassuccessfully established. Significantly reduced airway inflammation inasthmatic mice by treatment of dexamethasone and Mycobacterium phlei.Normal control lung tissue structure is clear, the performance of airwayinflammation.2) asthma group of CD4+of CD25+regulatory T cells accountof CD4+T-cell percentage was significantly lower than the control group（P<0.05）, the CD4+T cells of Th17cells the percentage significantly withhigher than normal control group (P <0.05); asthma to dexamethasonetreatment group, asthma Mycobacterium phlei mice treated with CD4+CD25+regulatory T cells accounted for the percentage of CD4+T cells thanasthmatic mice significantly increased (P <0.05), still below the normalcontrol group mice (P <0.05), the percentage of Th17cells accounts for CD4+T cells compared with asthmatic mice decreased significantly (P <0.05);still higher than the normal control group mice (P <0.05); difference betweenthe two drug treatment group was no statistical significance (P>0.05).3)asthmatic mice lavage fluid, serum IL-10levels compared with control groupdecreased significantly (P <0.05), IL-17levels compared with control groupwas significantly increased (P <0.05); dexamethasone treatment asthma group,the treatment of Mycobacterium phlei asthma group lavage fluid, serum IL-10levels than the asthmatic group increased (P <0.05), but with the normalcontrol group was still lower (P <0.05), while IL-17levels of the asthmagroup declined (P <0.05), but still compared with normal control group (P <0.05); was no significant difference between the groups of the two drugtreatment groups (P>0.05).4) each group of mice spleen mononuclear cells,CD4+T cell subsets of CD4+CD25+regulatory T cell percentage and thepercentage of Th17cells is negatively correlated; CD4+CD25+regulatory Tcell ratio of/of Th17and bronchoalveolar lavage fluid (BALF) and serumIL-10expression level was positively related to IL-17expression wasnegatively correlated; mice bronchoalveolar lavage fluid (BALF) IL-10, IL-17and serum IL-10, IL-17expression levels were positivelycorrelated.Conclusion:1). Asthmatic mice vivo existence of CD4+CD25+regulatory T cells in the number of reduction and IL-10generated lower, highexpression of of Th17cells and of IL-17to produce the increase and of CD4+of CD25+regulating sexual T cells/of Th17cells between the immuneimbalances in the immune dysfunction is one of the mechanisms of thepathogenesis of asthma.2).glucocorticoid increase of CD4+CD25+regulatory T cells and promote the generation of IL-10inhibit the expressionof Th17cells and IL-17production and correct the CD4+CD25+regulatoryT cells/Th17cells immune imbalance may be one of the mechanisms ofasthma treatment.3). Mycobacterium phlei after the intervention application asan immunomodulatory agent to play a similar role with glucocorticoids toimprove immunity, but its action and pathway needs further study. |
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