Neuroprotective effects of miR-30c on rats with cerebral ischemia/reperfusion injury by targeting SOX9

Pathol Res Pract. 2020 Dec;216(12):153271. doi: 10.1016/j.prp.2020.153271. Epub 2020 Nov 1.

Abstract

Objective: To explore the neuroprotective effect and mechanism of miR-30c in rats with cerebral ischemia/reperfusion (CI/R) injury.

Methods: in vivo, miR-30c mimic was transfected before the rats were treated with CI/R injury, and the neurological damage was evaluated. The expression of miR-30c was analyzed by RT-PCR.The area of cerebral infarction and pathological damage in CI/R rats were detected by TTC, H&E, Nissl and TUNEL staining. The expression of SOX9, BNDF and NT-3 were analyzed by immunofluorescence and western blot. The target relationship between SOX9 and miR-30c was analyzed by dual- luciferase reporter assay. in vitro, miR-30c mimic, SOX9 mimic and negative control were transfected before the OGD/R model of HT22 was established. CCK8 and flow cytometry were used to analyze the effect of miR-30c on the cell viability and apoptosis of HT22 cells in OGD/R condition. The protein expression of SOX9, BNDF and NT-3 were analyzed by immunofluorescence and western blot.

Results: The expression of miR-30c in the CI/R rats were significantly lower than that of sham group (p < 0.05). The area of cerebral infarction and the number of pathological damage and apoptosis in hippocampus were improved by miR-30c. miR-30c reduced SOX9 expression and improved BNDF and NT-3 expression in hippocampus compared with CI/R group (p < 0.05). The results of dual-luciferase reporter showed that SOX9 was the target of miR-30c. Under OGD/R condition, miR-30c mimic promoted cell viability and reduced cell apoptosis. miR-30c decreased the expression of SOX9, which was consistent with the results of immunofluorescence. Moreover, miR-30c could increase the expression of BNDF and NT-3 in HT22 cells. After transfection of SOX9 mimic, it was found that the above effects of miR-30c were all reversed.

Conclusion: miR-30c can improve the pathological damage of rats with CI/R injury and plays a neuroprotective role both in vivo and in vitro by targeting SOX9.

Keywords: Cerebral ischemia/reperfusion injury; Neuroprotection; SOX9; miR-30c.

MeSH terms

  • Animals
  • Apoptosis
  • Brain-Derived Neurotrophic Factor / metabolism
  • Cell Line
  • Cell Proliferation
  • Disease Models, Animal
  • Hippocampus / metabolism*
  • Hippocampus / pathology
  • Infarction, Middle Cerebral Artery / genetics
  • Infarction, Middle Cerebral Artery / metabolism
  • Infarction, Middle Cerebral Artery / pathology
  • Infarction, Middle Cerebral Artery / prevention & control*
  • Male
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Neurons / metabolism*
  • Neurons / pathology
  • Neurotrophin 3 / metabolism
  • Rats, Sprague-Dawley
  • Reperfusion Injury / genetics
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control*
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism*

Substances

  • Bdnf protein, rat
  • Brain-Derived Neurotrophic Factor
  • MIRN30 microRNA, rat
  • MicroRNAs
  • Neurotrophin 3
  • SOX9 Transcription Factor
  • Sox9 protein, rat