摘要
目的研究IL-17A在肺纤维化发病机制中的作用。方法 20只雌性Wistar大鼠,随机分为生理盐水(NS)组和博来霉素(BLM)组,NS组大鼠气管内灌注NS,BLM组大鼠气管内灌注BLM,两组分别于气管内灌注药物后第7天和第28天各处死一半动物。HE和Masson染色观察肺组织病理变化;免疫组织化检测肺组织IL-17A表达;收集支气管肺泡灌洗液(BALF),一部分行细胞数测定并进行分类分析,ELISA检测BALF中IL-17A的含量,另外一部分行分离、纯化得到肺泡巨噬细胞(AM),对其进行培养得到AM培养上清液(AMS),ELISA检测上清液IL-17A的含量,反转录PCR(RT-PCR)检测AM IL-17A mRNA表达。结果与NS组相比,BLM第7天组肺泡炎较明显,BALF中细胞总数增加,AM减少、中性粒细胞增加;第28天组肺泡炎减轻,而肺纤维化程度较重。与NS组比较,BLM第7天组和第28天组肺组织IL-17A表达明显增加(P<0.05),第28天较第7天的表达有所降低。与NS组比较,BLM组BALF中细胞总数第7天明显增高(P<0.05),第28天恢复至正常水平;BLM组BALF中IL-17A含量第7天和第28天明显增高,但第28天时较第7天降低;与NS组比较,BLM第7天组和第28天组AM培养前12 h、前24 h,前48 h上清液中IL-17A的含量均明显增加(P<0.05);RT-PCR结果显示,与NS组比较,BLM第7天组和第28天组各时间点AM IL-17A mRNA表达均明显增加(P<0.05)。结论 IL-17A促进了肺纤维化大鼠肺组织的炎症形成,进而参与了肺纤维化。
Objective To investigate the role of IL-17A in the development of pulmonary fibrosis in rats. Methods Twenty female Wistar rats were randomly divided into normal saline （NS） group and bleomycin （BLM） group. The BLM group was intratracheally instilled with BLM, while the NS group was treated with saline instead. Half of the rats in each group were sacrificed respectively on day 7 and 28 after intratracheal instillation. HE and Masson stainings were performed to show the pathological changes of lung tissues. The expression of IL-17A in lung tissues was detected by the immunohistochemistry. The bronchoalveolar lavage fluid （BALF） was harvested; one part of it was used for cell counting and classification, and for the detection of the concentration of IL-13A in BALF using ELISA; the other part of it was used for determining the concentration of IL-17A in the culture supematant of the purified alveolar macrophage （AM） using ELISA and reverse transcription PCR （RT-PCR）. Results Compared with the NS group, the BLM group presented a great deal of inflammatory cells in the alveoli, the increased total number of cells, decreased AM level and increased neutrophil number in BALF on day 7 ; on day 28, the BLM group had milder alveolitis and more serious fibrosis. The expression of IL-17A protein in lung tissues was obviously higher on day 7 and 28 in the BLM group than in the NS group （ P〈0.05）, and it was higher on day 7 than on day 28. The increased total number of BALF cells in BLM group returned to the normal level on day 28. The level of IL-17A in BALF increased significantly on day 7 and 28 in BLM group, however, compared with that on day 7, it decreased on day 28. Compared to the NS group, the expression of IL-17A protein in the culture supernatant of the BLM group was remarkably higher at 12th, 24th and 48th hours （ P 〈 0.05）. RT-PCR showed that compared with the NS group, the expression of IL-]TA mRNA increased significantly in BLM group on day ？ and 28 （ P 〈 0.05）. Conclusion IL-17A promotes the development of pulmonary inflammation, indicating that it may be involved in the progress of pulmonary fibrosis.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第4期366-370,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
泸州市科技局资助项目(12102)
