Metabolic Status and Morphological Changes of Astrocytes under Reductive Stress in vitro: Neural Network Analysis

Reductive stress caused by excessive accumulation of reducing equivalents (NADH, NADPH, glutathione), is increasingly recognized as a pathogenetic factor at the early stages of neurodegenerative diseases. However, its impact on the morphofunctional properties of astrocytes remains poorly understood. We performed the first comprehensive quantitative assessment of biochemical and morphological changes in cultured primary rat astrocytes under chronic reductive stress modeled using dithiothreitol (DTT, 500 μM, 24 h). To enable morphometric analysis of live, unstained cells, we developed and trained a semantic segmentation model based on the YOLOv11 architecture, which enables objective assessment of complex branched cell morphology in phase-contrast images while avoiding fixation-related artifacts. DTT-induced reductive stress caused a significant increase in the total pool of nicotinamide coenzymes (p < 0.01) and elevated mitochondrial superoxide production without compromising mitochondrial membrane potential. Morphometric analysis revealed a sustained enlargement of astrocyte soma area and increase in the number of astrocyte processes, along with a reduced branching complexity. These findings provide new insights into the role of redox imbalance in regulating glial function and may expand our understanding of the early mechanisms of neurodegenerative diseases.