天津医药 ›› 2017, Vol. 45 ›› Issue (1): 9-12.doi: 10.11958/20161020

• 细胞与分子生物学 • 上一篇    下一篇

不同程度间歇低氧对小鼠肺成纤维细胞活化和胞外基质分泌的影响 #br# #br#

李菲 1, 李莲 1, 任方元 1, 姜芳馨 , 冯靖 , 陈宝元 , 曹洁    

  1. 1. 天津医科大学总医院呼吸科
    2. 天津医科大学总医院
    3. 南开大学
  • 收稿日期:2016-09-22 修回日期:2016-11-12 出版日期:2017-01-15 发布日期:2017-01-15
  • 通讯作者: 曹洁 E-mail:tjcaojie@sina.com
  • 基金资助:
    国家自然科学基金;天津医科大学总医院青年孵育基金

Effects of different degrees of intermittent hypoxia on the activation and the extracellular matrix secretion of mouse lung fibroblast cells

  • Received:2016-09-22 Revised:2016-11-12 Published:2017-01-15 Online:2017-01-15
  • Contact: CAO Jie E-mail:tjcaojie@sina.com

摘要: 摘要: 目的 探讨不同程度间歇低氧 (IH) 对 MLg 小鼠肺成纤维细胞活化和胞外基质分泌的影响。方法 取对 数生长期的 MLg 成纤维细胞, IH 分别处理 1 h、 4 h、 8 h (IH1 组、 IH4 组、 IH8 组), IH 频率为 5% O2100 s, 21% O2 120 s, 并设置常氧对照组(21% O2处理 8 h, N 组)。实验结束后收集细胞, 采用 qRT-PCR 和 Western blot 检测各组 α 肌动 蛋白(α-SMA)、 Ⅰ型胶质蛋白(COL1) mRNA 和蛋白表达水平。结果 IH1 组、 IH4 组、 IH8 组 α-SMA、 COL1 mRNA 及蛋白的表达水平明显高于 N 组(P<0.05), 且随着 IH 时间的延长, α-SMA、 COL1 表达水平逐渐升高。结论 IH 可以促进小鼠肺成纤维细胞活化和胞外基质分泌, 其作用机制可能与氧化应激有关。

关键词: 缺氧, 成纤维细胞, 细胞外基质, 胶原Ⅰ型, 肌动蛋白类, 睡眠呼吸暂停, 阻塞性, 肺纤维化

Abstract: Abstract:Objective To explore the effects of different degrees of intermittent hypoxia (IH) on the activation and the secretion of extracellular matrix in MLg lung fibroblast cell line. Methods MLg lung fibroblast cells in logarithmic growth phase were exposed for 5% O2 for 100 seconds and 21% O2 for 120 seconds in 1 h, 4 h and 8 h groups (IH1, IH4 and IH8) and normoxia group (21% O2 for 8 h, N group). The cells in each group were collected at the end of experiment. Real-time PCR was used to measure the mRNA expression levels of α-SMA and type Ⅰ collagen (COL1) A1, and Western blot assay was used to detect the protein expression levels of α-SMA and COL1. Results The mRNA and protein expression levels of α- SMA and COL1 were significantly increased in IH1, IH4 and IH8 groups than those in N group (all P < 0.05). Furthermore, expression levels of α- SMA and COL1 showed a time- dependent increase with IH exposure time. Conclusion The intermittent hypoxia can promote the cell activation and the extracellular matrix secretion of mouse lung fibroblast cells, which may be related with the oxidative stress.

Key words: Key words: lung fibrosis, intermittent hypoxia, extracellular matrix, α-SMA, type I collagen