Related Experiment Video
Updated: Dec 29, 2025
Neuron-Macrophage Co-cultures to Activate Macrophages Secreting Molecular Factors with Neurite Outgrowth Activity
Published on: March 30, 2018
Brain-derived neurotrophic factor mediates macrophage migration inhibitory factor to protect neurons against
Su Hwan Bae1, Mi Ran Yoo1, Ye Yeong Kim2
1Department of Rehabilitation Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
Abstract:
Macrophage migration inhibitory factor (MIF) is a chemokine that plays an essential role in immune system function. Previous studies suggested that MIF protects neurons in ischemic conditions. However, few studies are reported on the role of MIF in neurological recovery after ischemic stroke. The purpose of this study is to identify the molecular mechanism of neuroprotection mediated by MIF. Human neuroblastoma cells were incubated in Dulbecco's modified Eagle's medium under oxygen-glucose deprivation (OGD) for 4 hours and then returned to normal aerobic environment for reperfusion (OGD/R). 30 ng/mL MIF recombinant (30 ng/mL) or ISO-1 (MIF antagonist; 50 μM) was administered to human neuroblastoma cells. Then cell cultures were assigned to one of four groups: control, OGD/R, OGD/R with MIF, OGD/R with ISO-1. Cell viability was analyzed using WST-1 assay. Expression levels of brain-derived neurotrophic factor (BDNF), microtubule-associated protein 2 (MAP2), Caspase-3, Bcl2, and Bax were detected by western blot assay and immunocytochemistry in each group to measure apoptotic activity. WST-1 assay results revealed that compared to the OGD/R group, cell survival rate was significantly higher in the OGD/R with MIF group and lower in the OGD/R with ISO-1 group. Western blot assay and immunocytochemistry results revealed that expression levels of BDNF, Bcl2, and MAP2 were significantly higher, and expression levels of Caspase-3 and Bax were significantly lower in the MIF group than in the OGD/R group. Expression levels of BDNF, Bcl2, and MAP2 were significantly lower, and expression levels of Caspase-3 and Bax were significantly higher in the ISO-1 group than in the OGD/R group. MIF administration promoted neuronal cell survival and induced high expression levels of BDNF, MAP2, and Bcl2 (anti-apoptosis) and low expression levels of Caspase-3 and Bax (pro-apoptosis) in an OGD/R model. These results suggest that MIF administration is effective for inducing expression of BDNF and leads to neuroprotection of neuronal cells against hypoxic injury.
Insights
Macrophage migration inhibitory factor (MIF) protects neurons from hypoxic injury. MIF administration boosts neuroprotective factors like BDNF and MAP2, enhancing neuronal survival after ischemic stroke.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Macrophage migration inhibitory factor (MIF) is crucial for immune function.
- Previous research suggests MIF offers neuroprotection in ischemic conditions.
- The specific mechanisms of MIF in post-ischemic stroke neurological recovery require further elucidation.
Purpose of the Study:
- To investigate the molecular mechanisms underlying MIF-mediated neuroprotection.
- To assess the impact of MIF on neuronal survival and apoptosis in an in vitro model of ischemic stroke.
Main Methods:
- Human neuroblastoma cells were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) to simulate ischemic stroke.
- Cells were treated with recombinant MIF or a MIF antagonist (ISO-1).
- Cell viability (WST-1 assay) and expression of key proteins (BDNF, MAP2, Caspase-3, Bcl2, Bax) were analyzed via Western blot and immunocytochemistry.
Main Results:
- MIF administration significantly increased neuronal cell survival compared to the OGD/R group.
- MIF treatment upregulated anti-apoptotic proteins (Bcl2) and neurotrophic factors (BDNF, MAP2) while downregulating pro-apoptotic proteins (Caspase-3, Bax).
- Conversely, the MIF antagonist ISO-1 reduced cell survival and altered protein expression towards apoptosis.
Conclusions:
- MIF administration confers neuroprotection in an in vitro model of ischemic injury.
- MIF promotes neuronal survival by upregulating BDNF and MAP2 expression and modulating apoptotic pathways.
- These findings highlight MIF's therapeutic potential for neurological recovery after ischemic stroke.
