摘要：目的  研究2-吲哚啉酮衍生物（PMID）对二氧化硅（SiO2）致小鼠肺纤维化的预防作用。方法  通过肺部喷雾给予SiO2制备小鼠肺纤维化模型，确定SiO2最佳剂量后造模，而后分别ig 给予PMID（5，15和20 mg·kg-1）或吡非尼酮（PFD）15 mg·kg-1，每天2次，连续28 d。采用HE染色检测肺组织病理损伤，Masson染色检测肺组织中胶原形成，免疫组化检测纤维化相关蛋白〔转化生长因子β（TGF-β）、Ⅰ型胶原A2（ColⅠA2）和α 平滑肌肌动蛋白（α-SMA）〕和炎症细胞标志物〔CD11b，F4/80 和CD3〕表达，实时PCR 检测肺组织中白细胞介素6（IL-6）mRNA 水平。结果  与正常对照组相比，模型组肺组织病理损伤严重，肺组织发生实变，炎症细胞浸润，炎症评分显著升高（P<0.01），且有大量的胶原物质积聚，蓝染部分>80%（P<0.01）；肺组织TGF-β，ColⅠA2 和α-SMA 蛋白表达增加（P<0.01），IL-6 mRNA 水平升高（P<0.01）。与模型组相比，PMID 5，15 和20 mg·kg-1组肺泡结构正常，组织病理损伤减轻（P<0.05），且仅有少量胶原物质积聚，蓝染部分<20%（P<0.05）；肺组织TGF-β，ColⅠA2 和α-SMA 蛋白表达显著降低（P<0.05，P<0.01），中性粒细胞和巨噬细胞数量明显降低（P<0.05），IL-6 mRNA 水平显著降低（P<0.01）。结论  PMID 可通过降低炎症反应有效预防SiO2所致小鼠肺纤维化。

关键词：2-吲哚啉酮衍生物；二氧化硅；肺纤维化；炎症细胞

中图分类号: R963       文献标志码: A

文章编号: 1000-3002-(2018)03-0183-09

DOI: 10.3867/j.issn.1000-3002.2018.03. 004

基金项目: 国家自然科学基金(31170712); 北京市科委G20工程医药产业创新研发课题(Z181100002218031)

作者简介: 刘婷，硕士研究生，主要从事肺纤维化相关药物的药理学及药代动力学研究。

通讯作者: 李长燕, Tel: 13683288672, E- mail：fmmli@163.com

收稿日期: 2018-02-26

Abstract: OBJECTIVE  To study the preventive effect of 3- (3- pyridylmethylidene)-2-indolinone(PMID) on pulmonary fibrosis induced by silica (SiO2) in mice. METHODS  A pulmonary fibrosis model of mice induced by SiO2 was established, and the model dosage of SiO2 was determined. After modeling, the mice were ig given sodium carboxymethyl cellulose (CMC-Na), PMID 5, 15 and 20 mg·kg-1 and pirfenidone (PFD) 15 mg·kg-1, twice daily for 28 d. HE staining was used to observe the pathological injury to lung tissues. Masson staining was used to analyze the formation of collagens in lung tissues. Immunohistochemical staining was performed to analyze the expression of fibrosis related proteins transforming growth factor-β (TGF-β), collagen alpha-2 typeⅠ(ColⅠA2), α-smooth muscle actin (α-SMA) and inflammatory markers, including CD11b, F4/80 and CD3. The mRNA expression of interleukin 6 (IL- 6) in lung tissues was analyzed by real-time PCR. RESULTS  Compared with the normal control group, pathological injuries to lung tissues of model group were severe, lung tissues were consolidated, inflammatory cells infiltrated, and the inflammatory score increased significantly (P<0.01). The model group had a large amount of collagen accumulation, the blue staining part was greater than 80% (P<0.01), and the lung tissue had high expressions of TGF-β, ColⅠA2 and α-SMA (P<0.01). At the same time, the mRNA expression level of IL-6 was higher than that of the normal control group (P<0.01). Compared with the model group, the alveolar structure in the PMID administration group was normal, and PMID significantly alleviated the histopathological damage (P<0.05). Only a small number of collagens were accumulated and the blue staining part was less than 20% (P<0.05). PMID could significantly decrease the expression of TGF-β, ColⅠA2 and α-SMA, and the number of neutrophils and macrophages in the PMID group decreased significantly (P<0.05), while the mRNA expression level of IL- 6 was significantly reduced (P<0.01). CONCLUSION PMID can effectively prevent the pulmonary fibrosis induced by SiO2 in mice.

Key words: 3-(3-pyridylmethylidene)-2-indolinone; silica; pulmonary fibrosis; inflammatory cell infiltration