天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (3): 234-238.doi: 10.11958/20161506

• 实验研究 • 上一篇    下一篇

实验性肺纤维化小鼠肺泡巨噬细胞凋亡的动态变化及意义

李鑫 1， 李琦 1， 2， 李祎 1， 苏程程 1， 周欣 3， 彭守春 1， 魏路清 1△， 姬文婕 1△   

  1. 1 中国人民武装警察部队后勤学院附属医院呼吸与重症医学科 （邮编 300162）； 2 锦州医科大学武警后勤学院附属医院研究生培养基地； 3 武警后勤学院附属医院心脏中心
  • 收稿日期:2016-12-09 修回日期:2017-01-23 出版日期:2017-03-15 发布日期:2017-03-21
  • 通讯作者: 魏路清， 姬文婕 E-mail:ji_wenjie@hotmail.com
  • 基金资助:
    国家自然科学基金资助项目 （81102088， 81441101， 81570335）； 天津市自然科学基金资助项目 （15JCZDJC35000）； 武警后勤学院附属医院重点项目 （FYZ201510， FYZ201605）

The dynamic change and significance of apoptosis on lung alveolar macrophages in pulmonary fibrosis model mice

LI Xin1, LI Qi1,2, LI Yi1, SU Cheng-cheng1, ZHOU Xin3, PENG Shou-chun1, WEI Lu-qing1△, JI Wen-jie1△   

  1. 1 Department of Respiratory and Critical Care Medicine, the Affiliated Hospital of Logistics University of Chinese Armed Police Forces, Tianjin 300162, China; 2 The Affiliated Hospital of Logistics University of Chinese Armed Police Forces， Postgraduate Training Base of Jinzhou Medical University; 3 Heart Center,the Affiliated Hospital of Logistics University of Chinese Armed Police Forces
  • Received:2016-12-09 Revised:2017-01-23 Published:2017-03-15 Online:2017-03-21
  • Contact: WEI Lu-qing, JI Wen-jie E-mail:ji_wenjie@hotmail.com

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摘要： 摘要： 目的 探讨博莱霉素诱导的肺纤维化模型小鼠肺泡巨噬细胞（AM）凋亡的动态变化及意义。方法 100只 C57BL/6 雄性小鼠随机分为对照组和模型组， 每组各 50 只。模型组采用口咽吸入法给予博莱霉素 A2 （2.5 mg/kg）建立肺纤维化模型， 对照组给予等体积生理盐水。建模后第 1、 3、 7、 14 和 21 天每组随机处死 10 只动物， 5 只采用苏木素-伊红（HE）染色观察肺组织病理变化， 计算炎症评分及肺系数； 另 5 只采用碱水解法测定肺组织羟脯氨酸（Hyp） 的含量； 记录肺泡灌洗液 （BALF） 中 AM 数目， 流式细胞术检测 AM 凋亡情况。结果 模型组小鼠第 3 天肺组织出现炎症细胞浸润、 肺泡壁水肿增厚， 炎症评分明显升高； 第 7 天时炎症程度继续加重， 炎症评分进一步升高； 第14 天和第 21 天时出现纤维增生和胶原沉积； 第 21 天时炎症评分较第 7 天和第 14 天下降。第 21 天时模型组 Hyp含量较第 1、 3、 7 天均明显升高， 但与第 14 天相比无明显差异。在整个实验过程中， 模型组的肺系数， AM 数目及凋亡率呈现先升高后降低的变化， 于第 7 天达到峰值。结论 在博莱霉素诱导的小鼠肺纤维化模型中， AM 的数目和凋亡水平在前期的炎症反应阶段明显升高， 在肺纤维化阶段出现下降。动态变化的 AM 可能在肺纤维化进展过程中发挥不同的作用。

关键词: 肺纤维化, 巨噬细胞, 肺泡, 羟脯氨酸, 细胞凋亡, 博莱霉素, 流式细胞术

Abstract: Abstract： Objective To observe the dynamic change and significance of apoptosis levels in the alveolar macrophages(AM) of the bleomycin-induced pulmonary fibrosis model mice. Methods A total of 100 mice were randomly divided into control group (n=50) and model group (n=50). The model group was received with bleomycin at 2.5 mg/kg via oropharyngeal instillation to molding. The control group was conducted with same volume of normal saline. Ten mice randomly selected were executed on the day 1, 3, 7, 14, and 21 after modeling in the two groups. Hematoxylin and eosin (HE) staining was used to observe the pathological changes of lung tissue and calculate the inflammation score and pulmonary coefficients in five mice of two groups. Alkali hydrolysis method was used to detect the content of hydroxyproline, and the bronchoalveolar lavage fluid (BALF) was collected to assess the cell counts and apoptotic levels of AM in another five mice. Results Mice of model group showed inflammatory cell infiltration and alveolar wall widening on day 3, along with an aggravate inflammation and increased inflammation score on day 7. The fibroplasia and collagen sediment were increased on day 14 and day 21. The inflammation score was decreased significantly in day 21 compared with those in day 7 and day 14. The hydroxyproline contents were increased significantly on day 21 than those on day 1, day 3 and day 7 in model group, but no significant difference when compared with those of day 14. The cell count and apoptotic rates of AM, and pulmonary coefficients showed changes of increasing first and then decreasing in model group, which reached the peak at day 7.Conclusion The cell counts and apoptotic levels increase in the preliminary stage of inflammatory and decrease in pulmonary fibrosis phase in bleomycin-induced pulmonary fibrosis model mice. These changes may play different roles in the progression of pulmonary fibrosis.

Key words: pulmonary fibrosis, macrophages, alveolar, hydroxyproline, apotosis, bleomycin, flow cytometry