基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1618-1623.

• 研究论文 • 上一篇    下一篇

脑缺血/再灌注损伤中产生IL-17A的神经细胞类型鉴定

刘婷1,韩松2,张颖1,李淑娟1,李俊发1   

  1. 1. 首都医科大学
    2. 首都医科大学神经生物学系
  • 收稿日期:2016-08-18 修回日期:2016-09-24 出版日期:2016-12-05 发布日期:2016-11-29
  • 通讯作者: 李淑娟 E-mail:shujuanli@ccmu.edu.cn
  • 基金资助:
    国家自然科学基金面上项目;国家自然科学基金面上项目;北京市自然科学基金重点项目

Identification of neural cell-type as source of IL-17A during cerebral ischemia/reperfusion injuries

  • Received:2016-08-18 Revised:2016-09-24 Online:2016-12-05 Published:2016-11-29
  • Supported by:
    the National Natural Science Foundation of China;the National Natural Science Foundation of China

摘要: 目的 在离体细胞水平鉴定脑缺血/再灌注损伤中产生白细胞介素-17A (IL-17A)的脑固有神经细胞类型。方法 利用原代培养小鼠脑皮层神经元、小胶质细胞和星形胶质细胞1~4 h氧-糖剥夺/24 h复糖复氧(OGD/R)模拟细胞离体缺血/再灌注损伤模型,用免疫荧光双标、实时定量聚合酶链式反应(RT-qPCR)和蛋白免疫印迹(Western blot)等技术,确定产生IL-17A的神经细胞类型。结果 三种神经细胞中,除NeuN阳性神经元外,小胶质细胞和星形胶质细胞在OGD/R处理后,均可表达IL-17A蛋白,且分别与其特定标志物Iba-1和GFAP存在共定位现象。星形胶质细胞经过1~6 h OGD/24 h R处理后,IL-17A mRNA表达水平随OGD时间延长显著增高,且在4 h OGD/R时达高峰[(2.74±2.48),P<0.001,每组n=5]。此外,OGD/R可促进星形胶质细胞表达IL-17A蛋白[(3.17±0.91),P<0.05,每组n=5]。结论 脑缺血/再灌注损伤中星形胶质细胞可能是产生IL-17A的主要来源。

关键词: IL-17A, 氧-糖剥夺/复糖复氧, 星形胶质细胞, 小胶质细胞, 神经元

Abstract: Objective To identify the neural cell-type as source of IL-17A during cerebral ischemia/reperfusion injuries in vitro. Methods Primary cultured mouse cortical neurons, microglia and astrocytes were subjected to 1~4 h oxygen-glucose deprivation/24h reoxygenation (OGD/R) simulating cell ischemia/reperfusion injury model in vitro, and then the double immunofluorescence staining with IL-17A and their specific markers of NeuN, Iba-1 and GFAP, RT-qPCR and Western blot were performed to determine the neural cell-type of IL-17A production. Results The primary cultured microglia and astrocytes (but not NeuN+ neurons) could express IL-17A and respectively co-localized with their specific markers Iba-1 and GFAP after OGD/R treatment. The mRNA expression levels of IL-17A increased with OGD duration and reached the peak at 4h OGD [(2.74±2.48),P<0.001, n=5 per group] in astrocytes after 1~6 h OGD/24 h R treatment. In addition, 4 h OGD/R treatment could promote IL-17A protein expression in primary cultured astrocytes [(3.17±0.91),P<0.05,n=5 per group]. Conclusions These results suggested that astrocytes might be the main source of IL-17A production during cerebral ischemia/reperfusion injuries.

Key words: IL-17A, OGD/R, Astrocytes, Microglia, Neurons

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