Oxidative stress due to increased reactive oxygen species (ROS) formation and/or inflammation is considered to play an important role in ischemic stroke injury. Leukemia inhibitory factor (LIF) has been shown to protect both oligodendrocytes and neurons from ischemia by upregulating endogenous anti-oxidants, though the effect of ischemia and the protective role of LIF treatment in mitochondrial function have not been studied. The goal of this study was to determine whether LIF protects ischemia-induced altered mitochondrial bioenergetics in reproductively senescent aged rats of both sexes (≥18 months old), approximately equivalent to the average age of human stroke patients. Animals were euthanized at 3 days after permanent middle cerebral artery occlusion (MCAO) surgery. We found that MCAO surgery significantly reduced mitochondrial oxidative phosphorylation in both the ipsilateral striatum and prefrontal cortex in male aged rats compared to their respective contralateral regions of the brain. MCAO injury showed mitochondrial bioenergetic dysfunction only in the striatum in female rats; however, the prefrontal cortex remained unaffected to the injury. LIF-treated rats significantly prevented mitochondrial dysfunction in the striatum in male rats compared to their vehicle-treated counterparts. Collectively, MCAO-induced mitochondrial dysfunction and LIF’s potential as a therapeutic biomolecule exhibited sex- and tissue-specific effects, varying between the striatum and prefrontal cortex in male and female rats.

Traumatic brain injury (TBI) triggers complex and evolving secondary cascades that disrupt mitochondrial homeostasis and contribute to progressive neurodegeneration. Although mitochondrial impairment is a well-recognized driver of post-traumatic pathology, the spatial and temporal progression of mitochondrial dysfunction, particularly in regions distal to the injury site, remains poorly defined, and potential sex-specific responses remain understudied. Here, we performed a comprehensive mitochondrial-focused analysis in a mouse model of controlled cortical impact (CCI), quantifying mtDNA copy number (mtDNA-CN), mitochondrial gene expression, and protein markers regulating biogenesis, transcription, electron transport chain integrity, and mitophagy. Mitochondrial profiles were assessed across four brain regions (cortex at 2, 4, and 6 mm from the injury epicenter, and hippocampus) at four time points (6h, 12h, 24h, and 48h) in both female and male C57BL/6J mice. While mtDNA content exhibited only modest and region-restricted reduction, particularly near the injury core, transcriptional and protein-level changes were far more pronounced and sex-divergent. Females displayed extensive early cortical gene activation followed by widespread hippocampal suppression at 48 h across mitochondrial dynamics, OXPHOS, transcriptional regulation, and biogenesis pathways, accompanied by 48h in PGC-1α, TFAM, and NDUFS1. In contrast, males showed minimal transcriptional disruption but demonstrated delayed compensatory increases in TFAM, NDUFS1, and p62 protein levels, suggesting activation of mitochondrial maintenance and recovery programs. These spatially and temporally distinct responses reveal fundamental sex-specific vulnerabilities in mitochondrial regulation after TBI. Together, our findings provide a direction to an integrated mitochondrial landscape of early post-injury events and identifies critical windows and pathways that may support sex-specific therapeutic targeting to restore mitochondrial function after TBI.

Stroke is a major cause of death and permanent disability, which is described by abrupt loss of neuronal energy, oxidative injury, inflammation, and apoptosis, primarily mediated by mitochondrial dysfunction. Mitochondria are key regulators of stress responses, apoptosis, redox homeostasis, immune signaling and known as central signaling hubs, integrating pathways from multiple cellular compartments to maintain homeostasis. Among the major regulatory elements are protein kinase enzymes that modulate cell signaling by phosphorylating substrates. Several kinases, including members of the Akt, PKA, PKC, GSK-3β, PINK1, and MAPK families, dynamically translocate to mitochondria under physiological and pathological conditions. Once localized, they influence mitochondrial dynamics, bioenergetics, reactive oxygen species (ROS) production, and programmed cell death. Dysregulation of these functions has been implicated in impaired mitophagy, aberrant calcium signaling, and processes associated with the pathogenesis of various neurological disorders, particularly in those with acute brain injuries, such as acute ischemic stroke (AIS). Especially, mitochondrial kinase oxidative stress hallmarks of neuronal injury. In this review, we examine the role of mitochondrial-associated kinases in AIS, explore mechanisms of their translocation, downstream signaling effects, and their promise as druggable targets highlighting the importance of spatial dynamics of kinases and the need for precision therapies. Understanding these mechanisms may open new avenues for therapeutic intervention in neurological diseases with a focus on acute brain injury, by targeting mitochondrial signaling networks.