Analysis of Caspase-9 protein and microRNAs miR-21, miR-126 and miR-155 related to the apoptosis mechanism in the cerebellum of rats submitted to focal cerebral ischemia associated with an alcoholism model

Arq Neuropsiquiatr. 2019 Oct 24;77(10):689-695. doi: 10.1590/0004-282X20190126. eCollection 2019.

Abstract

Objective: This study aimed to analyze the cerebellum of rats submitted to an experimental focal cerebral ischemia, by middle cerebral artery occlusion for 90 minutes, followed by reperfusion for 48 hours, associated with an alcoholism model.

Methods: Fifty adult Wistar rats were used, subdivided into five experimental groups: control group (C): animals submitted to anesthesia only; sham group (S): animals submitted to complete simulation of the surgical procedure; ischemic group (I): animals submitted to focal cerebral ischemia for 90 minutes followed by reperfusion for 48 hours; alcoholic group (A): animals that received daily absolute ethanol diluted 20% in water for four weeks; and, ischemic and alcoholic group (I + A): animals receiving the same treatment as group A and, after four weeks, submitted to focal cerebral ischemia for 90 minutes, followed by reperfusion for 48 hours. The cerebellum samples were collected and immunohistochemical analysis of Caspase-9 protein and serum analysis by RT-PCR of microRNAs miR-21, miR-126 and miR155 were performed.

Results: The expression of Caspase-9 was higher in groups I, A and I + A. In the microRNAs analyses, miR-126 was higher in groups A and I + A, miR-155 was higher in groups I and I + A.

Conclusions: We conclude that apoptosis occurs in the cerebellar cortex, even if it is distant from the ischemic focus, and that microRNAs 126 and 155 show a correlation with cellular apoptosis in ischemic rats and those submitted to the chronic alcohol model.

MeSH terms

  • Alcoholism / blood
  • Alcoholism / pathology*
  • Animals
  • Apoptosis*
  • Brain Ischemia / blood
  • Brain Ischemia / pathology*
  • Caspase 9 / analysis*
  • Cerebellum / chemistry
  • Cerebellum / pathology*
  • Immunohistochemistry
  • Infarction, Middle Cerebral Artery
  • Male
  • MicroRNAs / blood*
  • Random Allocation
  • Rats, Wistar
  • Real-Time Polymerase Chain Reaction
  • Reperfusion Injury / pathology
  • Time Factors

Substances

  • MicroRNAs
  • Caspase 9