Fructus xanthii Extract Alleviates Osteoarthritis by Preserving Cartilage Integrity

Osteoarthritis (OA) is a progressive degenerative joint disorder for which current pharmacological and surgical interventions mainly relieve symptoms rather than reverse the underlying pathological changes, highlighting the urgent need for novel therapeutic agents. This protocol aimed to investigate the potential chondroprotective effects of Fructus Xanthii extract against OA. First, non-targeted metabolomics profiling was conducted using ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) to characterize the chemical constituents of the extract. An integrated strategy combining network pharmacology and transcriptomic mining was then applied to identify bioactive components and putative targets of the extract, followed by cross-analysis with OA-related dysregulated genes to obtain core target genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to annotate the main biological processes and signaling pathways modulated by the extract. For in vivo validation, zebrafish cartilage injury models and mouse OA models were used to evaluate the chondroprotective effects of the extract. RNA sequencing (RNA-seq) was further adopted to analyze transcriptomic alterations in chondrocytes after treatment, focusing on OA-associated pathways and gene expression profiles; enzyme-linked immunosorbent assay (ELISA) was subsequently performed to verify the expression levels of key proteins/receptors screened by RNA-seq, so as to further confirm the regulatory effects of Fructus Xanthii extract on core OA targets. This integrated multi-omics and experimental protocol provides a systematic approach to explore the potential of Fructus Xanthii extract as a promising botanical candidate for OA intervention.