MiR-370 accelerated cerebral ischemia reperfusion injury via targeting SIRT6 and regulating Nrf2/ARE signal pathway

Kaohsiung J Med Sci. 2020 Sep;36(9):741-749. doi: 10.1002/kjm2.12219. Epub 2020 Apr 20.

Abstract

Cerebral ischemia reperfusion (CIR) is one of the highly lethal diseases in the world. MicroRNA-370 (miR-370) exerts multiple functions in different diseases. However, further research is needed to investigate the potential role of miR-370 in CIR injury. The in vivo middle cerebral artery occlusion (MCAO) rat model and in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) SH-SY5Y cell model were successfully established to mimic CIR injury. The infarct sizes of brain tissues from rats were evaluated. The relationship between miR-370 and silencing information regulatory protein 6 (SIRT6) was confirmed by luciferase activity assay. The cell viability and apoptosis were determined by CCK-8 assay and terminal-deoxynucleoitidyl transferase mediated nick end labeling staining. In this study, miR-370 was upregulated in brain tissues of MCAO rats and knockdown of miR-370 decreased cerebral infarction volume of MCAO rats and it alleviated CIR injury in vivo. The in vitro experiments indicated that knockdown of miR-370 promoted cell viability and alleviated OGD/R-induced SH-SY5Y cell apoptosis. Additionally, the TargetScan predicted that SIRT6 was a target of miR-370 and confirmed by luciferase activity assay. Moreover, miR-370 inhibited SIRT6 expression and regulated Nrf2/ARE signal pathway, whereas overexpression of SIRT6 partly reversed the effect of miR-370 on OGD/R-induced SH-SY5Y cell injury. Thus, we could conclude that miR-370 accelerated CIR injury via targeting SIRT6 and regulating Nrf2/ARE signal pathway, which might provide novel therapeutic targets for CIR injury treatment.

Keywords: MiR-370; Nrf2/ARE signal pathway; SIRT6; cerebral ischemia reperfusion injury.

MeSH terms

  • Animals
  • Base Pairing
  • Base Sequence
  • Brain / metabolism*
  • Brain / pathology
  • Brain Ischemia / genetics*
  • Brain Ischemia / metabolism
  • Brain Ischemia / pathology
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cell Line, Tumor
  • Gene Expression Regulation
  • Genes, Reporter
  • Glucose / deficiency
  • Glucose / pharmacology
  • Humans
  • Infarction, Middle Cerebral Artery / surgery
  • Luciferases / genetics
  • Luciferases / metabolism
  • Male
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • NF-E2-Related Factor 2 / genetics*
  • NF-E2-Related Factor 2 / metabolism
  • Neurons / drug effects
  • Neurons / metabolism
  • Neurons / pathology
  • Oxygen / pharmacology
  • Poly(ADP-ribose) Polymerases / genetics
  • Poly(ADP-ribose) Polymerases / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / genetics*
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / pathology
  • Signal Transduction
  • Sirtuins / genetics*
  • Sirtuins / metabolism

Substances

  • MIRN370 microRNA, rat
  • MicroRNAs
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, rat
  • Luciferases
  • Poly(ADP-ribose) Polymerases
  • Casp3 protein, rat
  • Caspase 3
  • Sirtuins
  • sirtuin 6, rat
  • Glucose
  • Oxygen