IL-17A抑制Th2细胞分化减轻哮喘小鼠气道嗜酸性粒细胞炎症

发布时间：2018-04-28 12:39

本文选题：支气管哮喘 + 嗜酸性粒细胞　；参考：《第三军医大学学报》2017年10期

【摘要】：目的研究IL-17A对哮喘小鼠Th2细胞分化及其相关炎症的作用。方法 24只C57BL/6J小鼠按随机数字表法分为对照组、哮喘组和IL-17A处理组(n=8)。哮喘组和IL-17A处理组予以卵清蛋白(ovalbumin,OVA)致敏及激发。每次雾化激发前1 h,IL-17A处理组给予重组小鼠IL-17A气道滴入。各步对照均予以生理盐水。末次激发后24 h处死小鼠,收集支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)行细胞总数及分类计数。ELISA检测BALF中IL-4、IL-5、IFN-γ、IL-17A的浓度。HE和PAS染色及半定量评分评估小鼠肺部病理变化。流式细胞术检测脾脏和支气管淋巴结Th细胞分化。免疫磁珠分选健康小鼠幼稚CD4~+T细胞,用Th2极化培养基体外培养,并给予IL-17A或等量PBS干预,检测Th2细胞的增殖、凋亡和分化。结果哮喘组较对照组,BALF中细胞总数、嗜酸性粒细胞数及其比例(P0.05)、IL-4、IL-5、IL-17A浓度均显著增高(P0.05),IFN-γ浓度显著下降(P0.05);支气管、血管周围炎症细胞浸润和杯状细胞化生明显加重(P0.01);脾脏和淋巴结Th2细胞分化比例显著增高(P0.05)。IL-17A处理组较哮喘组,BALF中的细胞总数[(26.00±5.43)×10~4/mL vs(58.40±26.93)×10~4/mL,P0.05]、嗜酸性粒细胞数[(8.04±1.98)×10~4/mL vs(31.95±12.28)×10~4/mL,P0.05]及其比例[(29.93±3.03)%vs(53.47±6.62)%,P0.01]显著降低,而中性粒细胞数及其比例无明显变化;BALF中Th2相关因子IL-4浓度[(9.86±2.77)pg/mL vs(28.13±4.62)pg/mL,P0.01]、IL-5浓度[(7.30±0.50)pg/mL vs(10.50±1.10)pg/mL,P0.01]均显著降低;支气管、血管周围炎症细胞浸润减轻,HE染色半定量评分降低[(2.00±0.51)vs(3.12±0.64),P0.05],杯状细胞化生减少[(0.80±0.45)vs(2.40±0.55),P0.01];脾脏[(2.24±0.44)%vs(4.82±1.83)%,P0.01]和淋巴结[(7.05±0.58)%vs(10.57±1.35)%,P0.05]中Th2细胞分化比例显著减少。极化培养的幼稚CD4~+T细胞,予IL-17A干预后,诱导分化的Th2细胞比例显著减少(P0.05),而增殖和凋亡无显著变化。结论 IL-17A有抑制Th2细胞分化,减轻哮喘小鼠气道嗜酸性粒细胞炎症的作用。
[Abstract]:Objective to study the effect of IL-17A on Th2 cell differentiation and related inflammation in asthmatic mice. Methods 24 C57BL/6J mice were randomly divided into control group, asthma group and IL-17A treated group. Asthma group and IL-17A group were sensitized and stimulated by ovalbumin ovalbumin OVA. One hour before each atomization, IL-17A group was treated with recombinant IL-17A. Each step control was given normal saline. The mice were killed 24 hours after the last challenge, and the bronchoalveolar lavage fluido BALFs were collected and counted. Elisa was used to detect the concentration of IL-4 and IL-5IL-5IFN- 纬 IL-17A in BALF. The lung pathological changes were evaluated by HE, PAS staining and semi-quantitative scoring. The differentiation of Th cells in spleen and bronchial lymph nodes was detected by flow cytometry. Immature CD4T cells of healthy mice were separated by immunomagnetic beads and cultured in vitro on Th2 polarization medium. The proliferation, apoptosis and differentiation of Th2 cells were detected by IL-17A or PBS intervention. Results compared with the control group, the total number of cells, the number and proportion of eosinophilic granulocytes in the asthma group were significantly higher than those in the control group. The concentration of IL-4, IL-5, IL-17A in the asthma group was significantly higher than that in the control group, and the concentration of IFN- 纬 in the bronchi was significantly decreased than that in the control group. The infiltration of perivascular inflammatory cells and goblet cell metaplasia increased significantly (P 0.01), the differentiation ratio of Th2 cells in spleen and lymph nodes was significantly higher than that in asthma group [26.00 卤5.43 脳 10~4/mL vs(58.40 卤26.93 脳 10 ~ 4% mL0.05], the number of eosinophil cells [8.04 卤1.98) 脳 10~4/mL vs(31.95 卤12.28) 脳 10 ~ (4) mL _ (0.05) and the proportion [29.93 卤3.03)%vs(53.47 卤6.62] decreased significantly. However, the number and proportion of neutrophils did not change significantly. The concentration of Th2 related factor IL-4 [9.86 卤2.77)pg/mL vs(28.13 卤4.62 渭 g / mLP0.01] IL-5 [7.30 卤0.50)pg/mL vs(10.50 卤1.10 g / mLP0.01] was significantly decreased in bronchi. The semi-quantitative score of HE staining was decreased [2.00 卤0.51)vs(3.12 卤0.64 P05], the metaplasia of goblet cells was decreased [0.80 卤0.45)vs(2.40 卤0.55 P0.01], the percentage of differentiation of Th2 cells in spleen [2.24 卤0.44)%vs(4.82 卤1.83] and lymph node (7.05 卤0.58)%vs(10.57 卤1.35P 0.05) was significantly decreased. After treated with IL-17A, the proportion of differentiated Th2 cells decreased significantly, but the proliferation and apoptosis were not significantly changed. Conclusion IL-17A can inhibit the differentiation of Th2 cells and alleviate the airway eosinophil inflammation in asthmatic mice.
【作者单位】：第三军医大学新桥医院全军呼吸内科研究所全军呼吸病研究重点实验室;
【基金】：国家自然科学基金面上项目(81270075)~~
【分类号】：R562.25

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