LncRNA PEG11as aggravates cerebral ischemia/reperfusion injury after ischemic stroke through miR-342-5p/PFN1 axis

Life Sci. 2023 Jan 15:313:121276. doi: 10.1016/j.lfs.2022.121276. Epub 2022 Dec 7.

Abstract

Aim: LncRNAs are highly expressed in the CNS and regulate pathophysiological processes. However, the potential role of lncRNAs inischemic stroke (IS) remains unknown. In this study, we investigated the functions and possible molecular mechanism of lncRNA paternal expressed gene 11 antisense (PEG11as) in this process.

Methods: Middle cerebral artery occlusion/reperfusion (MCAO/R) mice model and N2a cells model from oxygen-glucose deprivation/reoxygenation (OGD/R) were used to simulate cerebral I/R in vivo and in vitro. High-throughput sequencing (RNA-Seq) was used todetect differential expression of lncRNAs in cerebral I/R. QRT-PCR was used to detect the expression of PEG11as and miR-342-5p. Bioinformatics analysis, FISH, luciferase reporter assay, RIP, Western blot, and immunofluorescence were used to detect the interaction between PEG11as, miR-342-5p and PFN1. The effect on neuronal apoptosis was analyzed using loss-of-function combined with TUNEL, Hoechst, and caspase3 activity assays.

Key findings: 254 lncRNAs were differentially expressed in MCAO1h/R6h mice. Among them, PEG11as was significantly up-regulated. PEG11as down-regulated could markedly attenuate the brain infarct volume, alleviate neurological deficit in vivo, and effectively promote neuron survival, attenuate neuronal apoptosis both in vivo and in vitro. FISH assay discovered that PEG11as was mainly located in the cytoplasm. Furthermore, we demonstrated that PEG11as was able to bind miR-342-5p to inhibit miR-342-5p activity, whereas the down-regulated of miR-342-5p resulted in profilin 1 (PFN1) overexpression and thus promoting apoptosis.

Significance: This study suggests that PEG11as regulates neuronal apoptosis by miR-342-5p/PFN1 axis, which may contribute to our understanding of pathogenesis and provide a potential therapeutic option for cerebral I/R.

Keywords: Cerebral ischemic reperfusion; PEG11as; PFN1; RNA-Seq; miR-342-5p.

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Brain Ischemia* / genetics
  • Brain Ischemia* / metabolism
  • Glucose / metabolism
  • Infarction, Middle Cerebral Artery / metabolism
  • Ischemic Stroke*
  • Mice
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Profilins
  • RNA, Long Noncoding* / genetics
  • RNA, Long Noncoding* / metabolism
  • Reperfusion Injury* / metabolism
  • Stroke* / genetics

Substances

  • RNA, Long Noncoding
  • Profilins
  • MicroRNAs
  • Glucose