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目的:观察妊娠期哮喘小鼠模型中Th17及Treg细胞活性分布和表达变化,探讨Th17/Treg细胞免疫平衡在妊娠期哮喘小鼠发病中的作用机制。方法:24只6～8周龄,雌性BALB/c小鼠随机分为4组,每组6只。以OVA/Al(OH)3混悬液致敏与激发构建妊娠期哮喘组(AP组)和非妊娠期哮喘组(ANP组)小鼠模型;健康妊娠组(HP组)和健康非妊娠组(HNP组)小鼠致敏与激发均以只含Al(OH)3的生理盐水混悬液替代。光镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞计数及分类;HE染色评价小鼠气道炎症程度及肺组织病理学变化;流式细胞术测定小鼠外周血Th17及Treg细胞所占CD4+T细胞比例,计算Th17/Treg细胞比值;RT-PCR检测小鼠肺组织中Th17及Treg细胞相关因子表达。结果:妊娠期哮喘小鼠BALF中炎症细胞分类及计数、肺组织病理学表现和活体肺功能气道高反应性Penh值等客观指标证实,妊娠哮喘小鼠模型构建成功。外周血中,AP组CD4+IL-17+T细胞(Th17)显著升高(P<0.01);Foxp3+T细胞(Treg)显著降低(P<0.01);与HP,HNP组相比,存在Th17/Treg比例失衡(P<0.01)。AP组小鼠肺组织中IL-17、IL-23,IL-10 mRNA表达水平均显著高于HP组及HNP组(P<0.01);IFN-γmRNA则显著低于HP组及HNP组(P<0.01);ANP组各因子mRNA水平变化与AP组一致,组间无显著差异。结论:妊娠期哮喘小鼠存在细胞免疫功能失调;Th17、Treg细胞数量及免疫平衡状态发生改变,可能是妊娠期哮喘发病过程中一个重要的决定因素。
Objective: To observe the distribution and expression of Th17 and Treg cell activity in the mouse model of asthma during pregnancy, and to explore the mechanism of Th17 / Treg immune balance in the pathogenesis of gestational asthma in mice. Methods: Twenty-four 6-8 week old female BALB / c mice were randomly divided into 4 groups with 6 mice in each group. The mouse model of gestational asthma group (AP group) and non-pregnant asthma group (ANP group) were sensitized and challenged with OVA / Al (OH) 3 suspension. The healthy pregnant group (HP group) and healthy non-pregnant group (HNP group) mice were sensitized and challenged with Al (OH) 3 saline suspension only. The number and classification of cells in bronchoalveolar lavage fluid (BALF) were observed under light microscope. The degree of airway inflammation and pathological changes in lung tissue were evaluated by HE staining. The levels of Th17 and Treg cells in peripheral blood of mice were determined by flow cytometry Accounting for the proportion of CD4 + T cells, calculate the ratio of Th17 / Treg cells; RT-PCR detection of mouse lung tissue Th17 and Treg cells related factor expression. Results: The classification and counting of inflammatory cells in BALF, the histopathological features of lung tissue and the Penh value of airway hyperresponsiveness in living pulmonary function in pregnant mice showed that the model of pregnancy in asthma mice was successfully constructed. In peripheral blood, the levels of CD4 + IL-17 + T cells (Th17) were significantly increased (P <0.01) and Foxp3 + T cells (Tregs) were significantly decreased in AP group compared with those of HP and HNP groups Th17 / Treg ratio imbalance (P <0.01). The levels of IL-17, IL-23 and IL-10 mRNA in lung tissue of AP group were significantly higher than those in HP group and HNP group (P <0.01), and the levels of IFN-γmRNA in AP group were significantly lower than those in HP group and HNP group <0.01). The changes of mRNA and protein levels in ANP group were the same as those in AP group. There was no significant difference between the two groups. CONCLUSION: There is cellular immune dysfunction in asthmatic mice during pregnancy; the number of Th17 and Treg cells and immune balance are changed, which may be an important determinant in the pathogenesis of asthma during pregnancy.