To investigate the regulatory effects of hypoxia combined with inflammation on microglial energy metabolism and explore the underlying synergistic mechanisms in order to provide novel targets for the prevention and treatment of high-altitude cerebral edema.

BV2 microglial cells were categorized into four experimental groups: control (Con), hypoxia (Hy, 1% O₂), lipopolysaccharide (LPS, 0.1 μg/mL), and hypoxia combined with LPS (Hy-LPS). Cell viability was evaluated using the CCK-8 assay. The levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) in the supernatant were determined via the Griess method and ELISA, respectively. Transcriptomic profiling was conducted via RNA sequencing (RNA-seq), followed by bioinformatic analyses including identification of differentially expressed genes (DEGs), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and Gene Set Enrichment Analysis (GSEA).

Hy-LPS co-treatment significantly suppressed cell viability and synergistically promoted the release of NO and inflammatory cytokines compared to individual stimuli. Transcriptomic analysis revealed 2060 differentially expressed genes (DEGs) in the Hy-LPS group, including 912 up-regulated and 1148 down-regulated genes. These DEGs were prominently enriched in pathways associated with hypoxia response, inflammatory immunity, cell cycle regulation, and metabolic reprogramming, indicating a robust synergistic effect between hypoxia and inflammation at the transcriptional level.

This study sheds light on the molecular network through which hypoxia complicated with inflammation synergistically regulates microglial energy metabolism and inflammatory responses across multiple pathways, thereby providing potential targets for mechanistic research and the prevention of high-altitude cerebral edema.