ROS-Driven STAT1 S-Glutathionylation Sustains IFNγ Signaling and Pro-Inflammatory Microglial Polarization

Oxidative stress is a major driver of neuroinflammation, yet the molecular redox mechanisms that shape microglial activation remain incompletely defined. Among reversible redox modifications, protein S-glutathionylation has emerged as a key regulator of signaling cascades under conditions of elevated Reactive Oxygen Species (ROS). While IFNγ is known to activate STAT1 and promote a pro-inflammatory microglial phenotype, the contribution of oxidative stress to this process is poorly understood. Here, we investigated the interplay between ROS and STAT1 signaling in IFNγ-stimulated microglial cells. We demonstrate that ROS not only enhance STAT1 phosphorylation but also promote its S-glutathionylation, a modification that sustains STAT1 transcriptional activity. This dual regulation leads to prolonged expression of pro-inflammatory mediators, including iNOS, COX2, TNFα, and IL-6. Importantly, STAT1-deficient cells fail to mount these responses, confirming STAT1 as a central redox-sensitive hub in microglial polarization. Our findings identify S-glutathionylation as a molecular switch that links oxidative stress to persistent STAT1 activation and M1 polarization. These results suggest that targeting STAT1 redox regulation could help control microglial overactivation and may offer new opportunities for therapeutic intervention in neuroinflammatory and neurodegenerative diseases.