Effects and mechanism of CYBB on the progression of multiple sclerosis: Evidence from bioinformatic analysis and experimental validation.

BACKGROUND: Neutrophil extracellular traps (NETs) are involved in the pathological progression of multiple sclerosis (MS), yet the intrinsic molecular mechanism by which NETs mediate the progression of MS remains unclear. METHODS: Differentially expressed genes were screened by integrating Gene Expression Omnibus (GEO) datasets and NETs-related genes, and candidate biomarker genes for MS were identified using machine learning algorithms and receiver operating characteristic (ROC) curve analysis. Gene Set Enrichment Analysis (GSEA), immune infiltration analysis, construction of competitive endogenous RNA (ceRNA) networks and drug-gene interaction networks were performed on these candidate biomarker genes. An experimental autoimmune encephalomyelitis (EAE) mouse model was established and intervened with cytochrome b-245 beta chain (CYBB) overexpression. Neurological scores, inflammatory factors (tumor necrosis factor-α [TNF-α], interleukin-1β [IL-1β], interleukin-18 [IL-18], and monocyte chemoattractant protein-1 [MCP-1]), oxidative stress indicators (malondialdehyde [MDA], superoxide dismutase [SOD], and catalase [CAT]), pathological damage, and pyroptosis-related proteins (NOD-like receptor pyrin domain-containing 3 [NLRP3], apoptosis-associated speck-like protein containing a CARD [ASC], and Caspase-1) were evaluated. RESULTS: Four genes (CYBB, NF-kappa-B inhibitor alpha [NFKBIA], C-X-C motif chemokine receptor 4 [CXCR4], and IL1B) associated with MS were identified via bioinformatic analysis, among which CYBB exhibited the strongest immune-related correlation. The results of animal experiments revealed that EAE mice presented with weight loss, aggravated inflammation and oxidative stress, as well as elevated expression of pyroptosis-related proteins. CYBB overexpression significantly reversed these pathological alterations, suppressed proinflammatory cytokines and oxidative stress, and prevented the upregulation of pyroptosis-related proteins. CONCLUSION: CYBB overexpression alleviated inflammation and oxidative stress, suppressed the expression of pyroptosis-related proteins, and was associated with the amelioration of EAE pathological symptoms.