Effect of MicroRNA-126a-3p on Bone Marrow Mesenchymal Stem Cells Repairing Blood-brain Barrier and Nerve Injury after Intracerebral Hemorrhage

J Stroke Cerebrovasc Dis. 2020 May;29(5):104748. doi: 10.1016/j.jstrokecerebrovasdis.2020.104748. Epub 2020 Mar 9.

Abstract

Objective: Intracerebral hemorrhage (ICH) is a disease that threatens human health due to its high morbidity and mortality. On behalf of finding the better methods in the treatment of ICH, researchers pay more attention to a new technology which is finding effective genes to modify stem cells.

Methods: In this study, we isolated, cultured and identified bone marrow mesenchymal stem cells (MSCs) in vitro. Further, the MSCs (transfected with lentivirus expressing microRNA-126a-3p (miR-126)) were injected into the type Ⅶ collagenase-induced ICH rats to investigate the recovery effects of blood-brain barrier (BBB) and nerve damage in vivo.

Results: The MSCs surface marker molecules (CD29: 98.5%; CD90: 96.5%) were highly expressed, and the blood cell surface molecule was negatively expressed (CD45: 2%). Meanwhile, it was verified that miR-126 facilitated the differentiation of MSCs into vascular endothelial cells, owing to the rise of markers (CD31 and VE-cadherin). The modified neurological severity score, modified limb placing test score, brain water content and evans blue content were reduced after transplanted miR-126-modified MSCs. It was found that miR-126 accelerated the differentiation of MSCs into vascular endothelial cells via immunohistochemical staining in vivo. HE staining indicated the area of edema was obviously decreased compared with that in ICH + vector-MSCs group. MiR-126-modified MSCs alleviated the cell apoptosis in brain tissues by TUNEL assay. In addition, the mRNA and protein expression of protease activated receptor-1 and matrix metalloproteinase-9 were diminished, whilst the expression of zonula occludens-1 (ZO-1) and claudin-5 were enhanced in ICH+miR-126-MSCs group. Immunofluorescence assay revealed that miR-126-modified MSCs decreased the disruption of tight junction (ZO-1 and claudin-5).

Conclusions: All data illustrate that miR-126-modified MSCs repair BBB and nerve injury after ICH.

Keywords: Intracerebral hemorrhage; blood-brain barrier; bone marrow mesenchymal stem cells; microRNA-126a-3p; nerve injury.

MeSH terms

  • Animals
  • Apoptosis
  • Blood-Brain Barrier / metabolism*
  • Blood-Brain Barrier / pathology
  • Blood-Brain Barrier / physiopathology
  • Capillary Permeability*
  • Cell Differentiation
  • Cells, Cultured
  • Cerebral Hemorrhage / metabolism
  • Cerebral Hemorrhage / pathology
  • Cerebral Hemorrhage / physiopathology
  • Cerebral Hemorrhage / surgery*
  • Disease Models, Animal
  • Endothelial Progenitor Cells / metabolism
  • Endothelial Progenitor Cells / transplantation*
  • Gene Expression Regulation
  • Male
  • Mesenchymal Stem Cell Transplantation*
  • Mesenchymal Stem Cells* / metabolism
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Motor Activity
  • Nerve Regeneration*
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / transplantation*
  • Rats, Sprague-Dawley
  • Reflex
  • Sensation
  • Signal Transduction
  • Tight Junctions / metabolism
  • Tight Junctions / pathology

Substances

  • MicroRNAs