天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (2): 160-163.doi: 10.11958/20161305

• 实验研究 • 上一篇    下一篇

间歇低氧下小鼠血管内皮功能障碍机制的研究

王新 1， 孙蓓 2， 刘芳 1， 冯靖 1△， 曹洁 1△   

  1. 1 天津医科大学总医院呼吸科 （邮编 300052）； 2 天津医科大学代谢病医院内分泌研究所
  • 收稿日期:2016-11-08 修回日期:2016-11-30 出版日期:2017-02-15 发布日期:2017-02-14
  • 通讯作者: △通讯作者 冯靖 E-mail: zyyhxkfj@126.com； 曹洁 E-mail: tjcaojie@sina.com E-mail:zyyhxkfj@126.com
  • 作者简介:王新 （1990）， 男， 硕士在读， 主要从事睡眠低氧性疾病研究
  • 基金资助:
    国家自然科学基金资助项目 （81270144， 30800507， 81570084）

Studies of the mechanism of endothelial dysfunction in rats under intermittent hypoxia

WANG Xin1, SUN Bei2, LIU Fang1, FENG Jing1△，CAO Jie1△   

  1. 1 Department of Respiration, General Hospital of Tianjin Medical University, Tianjin 300052, China; 2 Institute of Endocrinology of Tianjin Medical University
  • Received:2016-11-08 Revised:2016-11-30 Published:2017-02-15 Online:2017-02-14
  • Contact: △Corresponding Author FENG Jing E-mail: zyyhxkfj@126.com; CAO Jie E-mail：tjcaojie@sina.com E-mail:zyyhxkfj@126.com

PDF (PC)

4019

摘要： 目的 探究间歇低氧 （IH） 条件下引起小鼠血管内皮功能障碍的机制。方法 呼吸仿真系统构建 IH 暴露模型， 将 60 只 C57BL/6J 雄性小鼠随机分为对照组和 IH 组。IH 组每天低氧暴露 8 h， 持续 6 周。应用酶联免疫吸附法 （ELISA） 测定小鼠血清低氧诱导因子-1a （HIF-1a） 和基质细胞衍生因子-1a （SDF-1a） 的水平； 测定 2 组小鼠血清中活性氧簇 （ROS） 的水平； 运用实时荧光定量 PCR 检测 2 组小鼠血清中 miR-199a-5p 的表达量； 应用双荧光素酶报告系统及点突变试验对 HIF-1a 进行靶基因验证。结果 与对照组相比， IH 组小鼠 HIF-1a（μg/L： 1.60±0.02 vs. 1.19±0.02）、 SDF-1a（ng/L： 1 823.00±8.97 vs. 1 444.00±17.90）的水平明显升高（P＜0.01）； ROS 水平明显高于对照组（U/mL： 487.66±35.73 vs. 211.57±23.82， P＜0.01）； IH 组的血清 miR-199a-5p 水平低于对照组（1.31±0.07 vs. 3.47± 0.17， P＜0.01）； 双荧光素酶报告基因检测结果证实 HIF-1a 为 miR-199a-5p 的靶基因。结论 IH 可引起小鼠血清 miR-199a-5p 水平的下降， 其靶基因 HIF-1a 继而升高， 而 HIF-1a 可引起 SDF-1a 升高， SDF-1a 的受体——细胞膜半胱氨酸-X-半胱氨酸受体-4 （CXCR4） 随之增加， 最终引起血清 ROS 水平升高， 导致血管内皮功能障碍。

关键词: 缺氧, 缺氧诱导因子 1, &alpha, 亚基, 受体, CXCR4, 微 RNAs, 活性氧, 间歇低氧, 基质细胞衍生因子-1a, miR-199a-5p

Abstract: Objective To explore the mechanism of vessel endothelial dysfunction in rats under intermittent hypoxia (IH). Methods The respiratory simulation system was used to simulate IH. Sixty C57BL/6J rats (male) were randomized into control group and IH group. The rats of IH group were exposed to IH 8 hours per day for 6 weeks. The serum levels of hypoxia inducible factor (HIF)-1a and stromal cell derived factor (SDF)-1a were assessed by ELISA. The serum levels of reactive oxygen species (ROS) were detected in two groups. The serum expression of miR- 199a-5p was detected by realtime fluorescent quantitative PCR in two groups. The dual luciferase report system and point mutation test were used to verify target gene for HIF-1a. Results The serum levels of HIF-1a and SDF-1a were significantly higher in IH group than those of control group (μg/L: 1.60±0.02 vs. 1.19±0.02, 1 823.00±8.97 vs. 1 444.00±17.90, P＜0.01). The serum level of ROS was significantly higher in IH group than that of control group (U/mL: 487.66±35.73 vs. 211.57±23.82, P＜0.01). The serum level of miR-199a-5p expression was significantly lower in IH group compared to that of control group (1.31±0.07 vs. 3.47± 0.17, P＜0.01). The result of dual luciferase reporter gene detection confirmed that target gene of miR-199a-5p was HIF- 1a. Conclusion The serum level of miR- 199a- 5p is decreased first due to IH, and then its target gene (HIF- 1a) is increased. HIF-1a can induce the increased level of SDF-1a, and its receptor (CXCR-4 ) is also increased. Finally, HIF- 1a can increase the serum level of ROS, resulting in the endothelial dysfunction.

Key words: anoxia, hypoxia- inducible factor 1, alpha subunit, receptors, CXCR4, microRNAs, reactive oxygen species, intermittent hypoxia, stromal cell derived factor-1a, miR-199a-5p