上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (5): 469-.doi: 10.3969/j.issn.1674-8115.2019.05.006

• 论著·基础研究 • 上一篇    下一篇

长链非编码 RNA Peg13在七氟烷所致的发育期原代神经元凋亡中的作用

蒋云凤，程燕咏，孙宇   

  1. 上海交通大学医学院附属第九人民医院麻醉科，上海 200011
  • 出版日期:2019-05-28 发布日期:2019-07-26
  • 通讯作者: 孙宇，电子信箱：dr_sunyu@163.com。
  • 作者简介:蒋云凤（1994—），女，硕士生；电子信箱： 15996931019@163.com。
  • 基金资助:
    上海市自然科学基金（18ZR1422900）

Role of long non-coding RNAPeg13 in apoptosis of developing primary neurons following sevoflurane injury

JIANG Yun-feng, CHENG Yan-yong, SUN Yu   

  1. Department of Anesthesiology, Shanghai Ninth Peoples Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
  • Online:2019-05-28 Published:2019-07-26
  • Supported by:
    Natural Science Foundation of Shanghai, 18ZR1422900

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摘要： 目的 ·研究长链非编码 RNA（long non-coding RNA，lncRNA）Peg13（paternally expressed 13）在七氟烷所致的发育期原代神经元凋亡中的作用。方法 ·以胎龄为 14.5 d的小鼠胎鼠的原代神经元为研究对象，实时定量 PCR检测七氟烷处理不同时间后 lncRNA Peg13的表达水平。荧光原位杂交实验检测 lncRNA Peg13在原代神经元中的定位。构建 lncRNA Peg13过表达和敲减质粒，分别转染原代神经元。4.1%的七氟烷处理 6 h后，荧光显微镜观察原代神经元的形态改变；采用 CCK-8分析原代神经元的活力；采用 TUNEL、 Western blotting检测原代神经元的凋亡程度。结果 ·在七氟烷处理的原代神经元中， lncRNA Peg13的表达水平呈时间依赖性下降。 LncRNA Peg13在原代神经元的细胞质和轴突中广泛表达。过表达 lncRNA Peg13可明显缓解七氟烷处理所致的神经元形态改变，导致神经元活力提高，凋亡细胞比例减少， Bcl-2、Bax蛋白表达量的比值（ Bcl-2/Bax）升高， caspase-3的表达水平降低。敲减 lncRNA Peg13可明显加剧七氟烷处理所致的神经元形态改变，导致神经元活力降低，凋亡细胞比例增加， Bcl-2/Bax降低， caspase-3的表达水平升高。结论 · LncRNA Peg13能够缓解七氟烷诱导的发育期原代神经元的凋亡。

关键词: 七氟烷, 长链非编码 RNA Peg13, 发育期原代神经元, 细胞活性, 细胞凋亡

Abstract:

Objective · To investigate the role of long non-coding RNA (lncRNA) Peg13 (paternally expressed 13) in the apoptosis of developing primary neurons following sevoflurane injury. Methods · Primary neurons were prepared fetal mice with 14.5 d of gestational age. The of lncRNA Peg13 after sevoflurane treatment was detectedquantitative PCR. The localization of lncRNA Peg13 in primary neurons was detectedin situ hybridization. Peg13 over and knockdown plasmids were constructed and transfected into primary neurons. The morphology of primary neurons was observedfluorescence microscope. Cell vability was assessedCCK-8 assay. Cell apoptosis was determinedTUNEL assay and Western blotting. Results · The of lncRNA Peg13 in primary neurons decreased in a time-dependent manner after sevoflurane treatment. LncRNA Peg13 was widely expressed in the cytoplasm and axon of primary neuron. Over of lncRNA Peg13 resulted in decreased sevoflurane-induced apoptosis. The primary neurons were restored to normal morphology with increased cell vability. The percentage of TUNEL-positive cells was decreased. The ratio of Bcl-2/Bax was increased and the of casepase-3 was decreased. However, knockdown of lncRNA Peg13 aggravated the sevoflurane-induced apoptosis. The primary neurons had visible morphological deterioration and decreased cell vability, with increased percentage of TUNEL-positive cells, decreased ratio of Bcl-2/Bax, and increased of casepase-3. Conclusion · LncRNA Peg13 may alleviate sevofluraneinduced apoptosis in developing primary neurons.

Key words: sevoflurane, long non-coding RNA Peg13, developing primary neuron, cell vability, apoptosis

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