Ischemic stroke is a leading contributor to death and disability worldwide, driving extensive research into pharmacological treatments beyond thrombolysis. Macrophage migration inhibitory factor (MIF), a cytokine, is implicated in several pathological conditions. In this study, we examined the effects of MIF on autophagy in astrocytes under the condition of chemical hypoxia. Primary astrocytes were treated with cobalt chloride, a well-known drug for inducing chemical hypoxia, followed by Western blot analyses to assess the HIF-1α, MIF, and LC3 protein levels along with a CCK assay. Additionally, cobalt chloride-treated astrocytes were co-treated with the MIF inhibitor ISO-1, and Western blot analyses were performed for MIF and LC3. Cell viability was evaluated using the CCK assay in astrocytes treated with cobalt chloride and ISO-1, with additional rapamycin treatment. Our results show that ISO-1 reduced LC3-II levels in astrocytes exposed to high concentrations of cobalt chloride (1000 μM) for 6 h. Moreover, rapamycin decreased cell viability in astrocytes treated with both 1000 μM cobalt chloride and ISO-1. Our data suggest that MIF plays a role in inducing autophagy in astrocytes under hypoxic conditions and is involved in the regulation of autophagic activity.
Keywords: MIF; autophagy; hypoxia; primary astrocytes.