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Published on: June 20, 2025
Hypoxia signaling regulates macrophage migration inhibitory factor (MIF) expression in stroke
Odysseus Zis1, Si Zhang, Katerina Dorovini-Zis
1Townsend Family Laboratories, Department of Psychiatry, Brain Research Center, Graduate Program in Neuroscience, The University of British Columbia, 2255 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada.
Abstract:
The macrophage migration inhibitory factor (MIF) is a multifaceted cytokine involved in many processes, including cellular responses to ischemia/reperfusion injury in the heart and brain. This study was undertaken to determine whether human MIF expression is induced following cerebral ischemia and its role therein. To examine whether the induction of MIF gene expression was mediated by its transcriptional upregulation, the human MIF gene promoter was cloned and a luciferase assay was used to determine the presence of a hypoxia-responsive region in the human MIF promoter. We found that human MIF promoter activity was significantly upregulated by hypoxia. A functional hypoxia-inducible factor 1α-binding site was identified using an electrophoretic mobility shift assay (EMSA). MIF has a protective effect on cortical neurons under oxygen-glucose deprivation (OGD) treatment. MIF significantly reduced OGD-induced cell death. To determine whether the expression of MIF in the human brain is altered following ischemia, brain sections from 10 stroke patients were examined with an antibody against MIF. Blood vessel endothelial cells in the peri-infarct region of ischemic brain displayed strong MIF immunoreactivity with no MIF immunoreactivity in control brains. Furthermore, we found that treatment of human brain endothelial cells with MIF had no effect on human monocyte adhesion to endothelium. Our study demonstrates that MIF gene expression is altered during stroke and dysregulation of the hypoxia signaling-induced MIF expression plays an important role in neuronal death in stroke.
Insights
Macrophage migration inhibitory factor (MIF) expression increases in the brain after stroke, offering neuroprotection. Dysregulation of hypoxia-induced MIF contributes to neuronal death in stroke patients.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Macrophage migration inhibitory factor (MIF) is a cytokine implicated in cellular responses to ischemia/reperfusion injury.
- Cerebral ischemia, a hallmark of stroke, involves complex cellular and molecular changes.
Purpose of the Study:
- To investigate whether human MIF expression is induced following cerebral ischemia.
- To elucidate the role of MIF in the context of stroke and its underlying molecular mechanisms.
Main Methods:
- Cloning and functional analysis of the human MIF gene promoter using luciferase assays.
- Identification of hypoxia-inducible factor 1α (HIF-1α) binding sites via electrophoretic mobility shift assays (EMSA).
- Assessment of MIF's neuroprotective effects on cortical neurons subjected to oxygen-glucose deprivation (OGD).
- Immunohistochemical analysis of MIF expression in brain tissue from stroke patients.
Main Results:
- Hypoxia significantly upregulated human MIF promoter activity, indicating transcriptional regulation.
- A functional HIF-1α binding site was identified within the human MIF promoter.
- MIF demonstrated significant neuroprotective effects, reducing OGD-induced neuronal cell death.
- Elevated MIF immunoreactivity was observed in endothelial cells of the peri-infarct region in stroke patient brains.
- MIF treatment did not affect monocyte adhesion to human brain endothelial cells.
Conclusions:
- Human MIF gene expression is altered in the brain following ischemic stroke.
- Hypoxia-induced MIF expression is transcriptionally regulated and plays a role in stroke pathophysiology.
- Dysregulation of hypoxia signaling-induced MIF contributes to neuronal death in stroke.
