To explore the potential targets and mechanisms of β-carotene in the treatment of hyperuricemia (HUA) by integrating network pharmacology analysis with in vivo mouse experiments.

Targets related to β-carotene were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), CTDbase, and Swiss Target Prediction databases. Therapeutic targets for HUA were obtained from the GeneCards, DigSee, and DisGeNET databases. Venn diagram analysis was used to identify overlapping targets between β-carotene-related targets and HUA disease targets, and a protein-protein interaction (PPI) network was constructed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the identified overlapping targets. C57BL/6 mice were divided into 5 groups: normal group, HUA (model group), positive control allopurinol (62.5 mg/kg) group, 20 mg/kg β-carotene group, and 40 mg/kg β-carotene group. Except for the control group, all other groups were induced to establish the HUA mouse model using potassium oxonate and hypoxanthine. Serum levels of uric acid (UA), urea nitrogen (BUN), and creatinine (CRE) were measured using biochemical assay kits. Renal tissue injury was observed using hematoxylin-eosin (H&E) staining.Levels of interleukin-1β (IL-1β) and IL-18 were detected using enzyme-linked immunosorbent assay (ELISA) kits. Western blot was used to detect protein expression levels of NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), and cleaved Caspase-1. Immunohistochemical staining was used to observe NLRP3 expression levels in renal tissue.

Matching 254 β-carotene-related targets with 170 HUA-related targets yielded 15 overlapping β-carotene-HUA target genes. KEGG enrichment indicated that β-carotene primarily treated HUA by involving inflammation and apoptosis pathways, with NLRP3 inflammasome enrichment being the most significant. Compared to the model group, both 20 mg/kg β-carotene and 40 mg/kg β-carotene significantly reduced serum UA (F=119.3, P<0.01), BUN (F=112.7, P<0.01), and CRE (F=80.76, P<0.01) levels in HUA mice, alleviated renal injury, decreased renal tissue levels of IL-1β (F=97.8, P<0.01) and IL-18(F=137.0, P<0.01), and downregulated the expression levels of NLRP3 (F=178.4, P<0.01), ASC (F=392.5, P<0.01), and cleaved-Caspase-1 (F=171.6, P<0.01).

β-Carotene can alleviate renal injury and reduce inflammatory responses in HUA mice. Its mechanism may be associated with inhibiting the activation of the NLRP3 inflammasome.