Key Regulatory Networks of Mitochondrial Dysfunction in Human Hippocampal Neurons under Cobaltous Chloride-Induced Hypoxia Revealed by Combined Transcriptome Analysis with Protein-Protein Interaction

Here, we exposed human hippocampal neuronal cells (HPPNCs) to CoCl₂-induced hypoxia and performed transcriptome sequencing alongside morphological and functional assays. We identified substantial mitochondrial damage and significant depolarization of mitochondrial membrane potential under hypoxic conditions.

RNA-seq analysis revealed 471 differentially expressed genes (DEGs) enriched in pathways related to oxygen sensing, metabolic reprogramming, angiogenesis, and synaptic function. Protein-protein interaction analysis, molecular docking, and ceRNA network construction highlighted HSPD1, HSPA9, and HSP90AA1 as central regulators of mitochondrial quality control and adaptation to hypoxic stress. For transcriptome analysis, total RNA was extracted, polyA-selected, and strand-specific libraries with ~400 bp inserts were prepared.