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目的观察苯亚甲基丙二腈脂类衍生物AG490对中性粒细胞性哮喘模型小鼠气道炎症的影响。方法无特定病原体(SPF)级C57BL雌性小鼠54只,按随机数字表法分成中性粒细胞性哮喘(NA)组、AG490处理的NA(NAAG)组和正常(NC)组,每组18只。NA、NAAG组小鼠于第0、6、13天经气道滴卵清蛋白(OVA)、脂多糖(LPS)致敏。NAAG组小鼠于实验第0天起经腹腔注射AG490每只500μg,3次/周,连续3周。于第21天连续2 d雾化吸入10 g/L OVA激发,每次1 h。最后1次雾化结束后24 h时,收集小鼠标本并检测相关指标。收集支气管肺泡灌洗液(BALF)并检测其有核细胞总数及分类比例;分离肺组织行HE染色及过碘酸希夫(PAS)染色,光镜下观察肺组织病理变化和杯状细胞增生;采用流式细胞术检测肺脏Th17细胞和调节性T细胞(Treg)的频数;采用ELISA检测小鼠BALF中IL-17的水平。结果与NA组相比,NAAG组BALF有核细胞计数、中性粒细胞百分比、嗜酸性粒细胞百分比均明显降低,NAAG组肺组织病理学改变及杯状细胞增生较NA组明显减轻。与NA组相比,NAAG组BALF IL-17水平降低、肺组织Th17细胞比例减少、Treg比例增加。结论 NA小鼠致敏阶段给予AG490处理,可增加小鼠激发阶段肺组织Treg数量、减少Th17细胞数量,减轻NA小鼠气道炎症。
Objective To observe the effect of benzalmalononitrile lipid derivative AG490 on airway inflammation in neutrophilic asthmatic mice. Methods Fifty-four female mice without specific pathogen (SPF) C57BL were divided into three groups according to random number table: neutrophil asthma (NA) group, AG490-treated NA group and normal group only. NA and NAAG mice were sensitized by OVA and LPS on days 0, 6 and 13. The mice in NAAG group were injected intraperitoneally with 500μg of AG490 on the 0th day of the experiment for 3 times / week for 3 weeks. On the 21st day for 2 consecutive days of inhalation of 10 g / L OVA challenge, each 1 h. Twenty-four hours after the last nebulization, the mouse specimens were collected and tested for relevant indicators. The bronchoalveolar lavage fluid (BALF) was collected and the total number of nucleated cells and the percentage of the sorted nuclei were detected. The lung tissues were isolated and stained with HE staining and periodic acid Schiff (PAS) staining. The pathological changes of lung tissue and goblet cell proliferation The frequency of Th17 cells and Tregs in lungs were detected by flow cytometry. The levels of IL-17 in BALF were detected by ELISA. Results Compared with NA group, the number of BALF nucleated cells, the percentage of neutrophils and the percentage of eosinophils in NAAG group were significantly decreased. The pathological changes of lung tissue and the goblet cell proliferation in NAAG group were significantly reduced compared with NA group. Compared with NA group, BALF IL-17 level was decreased in NAAG group, the proportion of Th17 cells in lung tissue decreased, and the proportion of Treg increased. Conclusion AG490 treatment in NA mouse sensitized phase can increase the number of Tregs in the lung tissue of the mice during the excitation phase, decrease the number of Th17 cells and reduce the airway inflammation in NA mice.