中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (11): 1919-1926.doi: 10.4103/1673-5374.238717

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

脑微血管内皮细胞损伤修复与长非编码RNA Snhg12及靶向miR-199a的调控机制

  

  • 收稿日期:2018-07-20 出版日期:2018-11-15 发布日期:2018-11-15
  • 基金资助:

    海南省自然科学基金（817334）

LncRNA SNHG12 ameliorates brain microvascular endothelial cell injury by targeting miR-199a

Fa-Qing Long, Qing-Jie Su, Jing-Xia Zhou, De-Sheng Wang, Peng-Xiang Li, Chao-Sheng Zeng, Yi Cai   

  1. The Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan Province, China
  • Received:2018-07-20 Online:2018-11-15 Published:2018-11-15
  • Contact: Yi Cai, caiyi@protonmail.com
  • Supported by:

    This study was supported by the Natural Science Foundation of Hainan Province of China, No. 817334.

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摘要：

最近大量研究报道长链非编码RNA（long non-coding RNAs，lncRNAs）能够调控缺血再灌注(ischemia/reperfusion；I/R)和氧糖剥夺/复氧（oxygen-glucose deprivation/reoxygenation, OGD/R）损害的脑微血管内皮细胞死亡、炎症反应以及血管生成，其在损害后的小鼠脑微血管内皮细胞中表达上调，但它是否与缺血性脑卒中有关尚不清楚。我们假设Snhg12在缺血性脑卒中有正向调控作用，但其作用机制又是什么呢？首先将微血管内皮细胞分别置于OGD环境中0，2，4，8，16和24 h，复氧4 h构建细胞的OGD/R模型，采用qRT-PCR检测的Snhg12 mRNA在脑微血管内皮细胞中的表达随OGD暴露时间的增加而上调，然后通过转染过表达或敲除脑微血管内皮细胞中的Snhg12，并将其暴露于OGD中16 h。采用MTT实验，锥虫蓝排斥实验，蛋白免疫实验和类毛细血管形成实验检测Snhg12在遭遇OGD/R损伤过程中对细胞生存、炎症，以及损伤后血管生成的影响发现，在遭遇OGD/R损伤过程中，Snhg12过表达可显著抑制脑微血管内皮细胞的死亡和炎症反应，并促进OGD/R损伤后的血管生成，而Sngh12敲除可呈现相反的作用。进一步观察到的分子机制是miR-199a被识别和证实能够与Snhg12相互作用，并且Snhg12过表达废除了miR-199a对细胞死亡、炎症反应以及血管生成的影响。上述数据证实了长链非编码RNA Snhg12通过靶向miR-199a抑制了由OGD/R引起的脑微血管内皮细胞的损伤。

orcid:0000-0001-6138-3349(Yi Cai)

 

关键词: 缺血性脑卒中, 长链非编码RNA, Snhg12, microRNA-199a, 脑微血管内皮细胞死亡, 炎症反应, 血管生成, 氧糖剥夺/复氧, 缺血/再灌注, 治疗靶点, 神经再生

Abstract:

Long non-coding RNAs regulate brain microvascular endothelial cell death, the inflammatory response and angiogenesis during and after ischemia/reperfusion and oxygen-glucose deprivation/reoxygenation (OGD/R) insults. The long non-coding RNA, SNHG12, is upregulated after ischemia/reperfusion and OGD/R in microvascular endothelial cells of the mouse brain. However, its role in ischemic stroke has not been studied. We hypothesized that SNHG12 positively regulates ischemic stroke, and therefore we investigated its mechanism of action. We established an OGD/R mouse cell model to mimic ischemic stroke by exposing brain microvascular endothelial cells to OGD for 0, 2, 4, 8, 16 or 24 hours and reoxygenation for 4 hours. Quantitative real-time polymerase chain reaction showed that SNHG12 levels in brain microvascular endothelial cells increased with respect to OGD exposure time. Brain microvascular endothelial cells were transfected with pcDNA-control, pcDNA-SNHG12, si-control, or si-SNHG12. After exposure to OGD for 16 hours, these cells were then analyzed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, trypan blue exclusion, western blot, and capillary-like tube formation assays. Overexpression of SNHG12 inhibited brain microvascular endothelial cell death and the inflammatory response but promoted angiogenesis after OGD/R, while SNHG12 knockdown had the opposite effects. miR-199a was identified as a target of SNHG12, and SNHG12 overexpression reversed the effect of miR-199a on brain microvascular endothelial cell death, the inflammatory response, and angiogenesis. These findings suggest that SNHG12 suppresses endothelial cell injury induced by OGD/R by targeting miR-199a.

Key words: nerve regeneration, ischemic stroke, microRNA, brain microvascular endothelial cell death, inflammatory response, angiogenesis, oxygen-glucose deprivation/reoxygenation, ischemia/reperfusion, therapeutic targets, neural regeneration, gene regulation, neural regeneration