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Author Spotlight: Establishing a Reliable Distal MCA Occlusion Model in Mice for Stroke Research
Published on: December 15, 2023
Regulatory effects of miR-146a/b on the function of endothelial progenitor cells in acute ischemic stroke in mice
Zeng-Feng Su1, Zhong-Wu Sun1, Ying Zhang2
1Department of Neurology, First Affiliated Hospital of Anhui Medical University, Hefei, PR China.
Abstract:
The study aims to explore how microRNA-146a/b (miR-146a/b) regulates the function of endothelial progenitor cells (EPCs) in acute ischemic stroke in mice. Eighty male SPF C57BL/6J mice were evenly divided into the model-6 h, model-12 h, model-24 h (mice suffered from middle cerebral artery occlusion [MCAO] for 6 h, 12 h and model-24 h) and normal groups. EPCs were transfected and assigned into the control, MCAO, MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. The qRT-PCR was used to detect miR-146a/b expression in EPCs. Expressions of tumor necrosis factor receptor-associated factor 6 (TRAF6) and interleukin-1 receptor-associated kinase 1 (IRAK1) were detected using western blotting. Cell proliferation and migration of EPCs were testified using CCK-8 assay and scratch test, respectively. Angiogenesis ability of EPCs was observed under microscope. MiR-146a and miR-146b expressions were lower in the model groups than the normal group. There were up-regulated TRAF6 and IRAK1 expressions in the model-6 h, model-12 h and model-24 h groups compared with the normal group. And there were down-regulated TRAF6 and IRAK1 expressions in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups than in the MCAO group. Compared with the control group, the proliferation, migration and angiogenesis ability of EPCs were significantly lower in the MCAO group, but higher in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. Besides, the miR-146a/b group showed more enhancement than the MCAO-miR-146a and MCAO-miR-146b groups. MiR-146a/b could down-regulate the TRAF6 and IRAK1 expressions and promote proliferation, migration and angiogenesis ability of EPCs, which was important for recovery of patients with hyperacute ischemic stroke.
Insights
MicroRNA-146a/b (miR-146a/b) enhances endothelial progenitor cell (EPC) function in acute ischemic stroke by down-regulating TRAF6 and IRAK1. This promotes EPC proliferation, migration, and angiogenesis, aiding recovery in hyperacute stroke patients.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Acute ischemic stroke is a leading cause of disability.
- Endothelial progenitor cells (EPCs) play a crucial role in vascular repair after stroke.
- MicroRNAs (miRNAs) are emerging as key regulators of cellular function in stroke pathogenesis.
Purpose of the Study:
- To investigate the role of microRNA-146a/b (miR-146a/b) in regulating EPC function in a mouse model of acute ischemic stroke.
- To elucidate the molecular mechanisms by which miR-146a/b affects EPCs, focusing on TRAF6 and IRAK1.
- To assess the therapeutic potential of miR-146a/b in promoting recovery from ischemic stroke.
Main Methods:
- Middle cerebral artery occlusion (MCAO) model in C57BL/6J mice.
- Quantitative real-time PCR (qRT-PCR) to measure miR-146a/b expression.
- Western blotting to detect TRAF6 and IRAK1 protein levels.
- Cell Counting Kit-8 (CCK-8) assay for cell proliferation.
- Scratch test for cell migration.
- Microscopic observation of angiogenesis.
Main Results:
- miR-146a/b expression was significantly reduced in EPCs from MCAO mice compared to normal controls.
- TRAF6 and IRAK1 expressions were upregulated in MCAO mice, and downregulated upon miR-146a/b transfection.
- EPC proliferation, migration, and angiogenesis were impaired in MCAO mice but significantly enhanced following miR-146a/b transfection.
- Co-transfection of miR-146a and miR-146b showed a greater restorative effect than individual miRNA transfection.
Conclusions:
- miR-146a/b plays a protective role in acute ischemic stroke by enhancing EPC function.
- miR-146a/b exerts its effects by down-regulating TRAF6 and IRAK1, thereby promoting EPC proliferation, migration, and angiogenesis.
- Restoring miR-146a/b levels represents a potential therapeutic strategy for improving recovery in hyperacute ischemic stroke.
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