These findings indicated that baicalin alleviated chondrocyte injury.

Exploration of molecular mechanism of baicalin targeting TRIM24 to suppress NLRP3/caspase-1-mediated chondrocyte pyroptosis in osteoarthritis based on network pharmacology and molecular docking.

Guanjian Liu, Hao Jiang, Fujun Dai, W Y Zhang, Shufan Ge, Jingwen Liu

Int Immunopharmacol

Dec 10, 2025

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Abstract

BACKGROUND: Baicalin is a major active ingredient from Scutellaria baicalensis with important roles in various inflammatory diseases, including osteoarthritis (OA). This study aimed to investigate whether baicalin regulates chondrocyte pyroptosis in OA by targeting tripartite motif-containing 24 (TRIM24). METHODS: Network pharmacological analysis was conducted to screen targets of baicalin and OA, followed by protein-protein interaction (PPI) network construction and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. For establishment of in vitro OA model, C28/I2 cells were treated with lipopolysaccharide (LPS) and adenosine triphosphate (ATP). Cell viability, cytotoxicity, and apoptosis were detected by CCK-8 assay, lactate dehydrogenase cytotoxicity assay, and flow cytometry/TUNEL assay, respectively. Markers of oxidative stress were measured using commercial kits. The levels of inflammatory cytokines were evaluated through enzyme-linked immunosorbent assay. The protein expression levels were examined by Western blotting. Caspase-1 activity was determined via caspase-1 activity assay. AutoDock vina software was used for molecular docking between baicalin and TRIM24. Additionally, baicalin and TRIM24 binding was validated using cellular thermal shift assay (CETSA). In vivo OA model was established by destabilized medial meniscus (DMM) surgery on Sprague-Dawley (SD) rats. RESULTS: A total of 201 intersecting genes between baicalin and OA were screened, including TRIM24. NOD-like receptor signaling pathway was found as one of the top 10 enriched pathways in the KEGG analysis. Baicalin mitigated LPS/ATP-induced cell viability inhibition, cytotoxicity and apoptosis in C28/I2 cells. Baicalin restrained pyroptosis in LPS/ATP-treated C28/I2 cells via obstructing the NLRP3/caspase-1 pathway. Baicalin interacted with TRIM24 and up-regulated TRIM24 protein expression. Baicalin protected C28/I2 cells by targeting TRIM24. TRIM24 could enhance NLRP3 ubiquitination and its downregulation reversed the anti-pyroptotic effect of baicalin via mediating the NLRP3/caspase-1 pathway. Baicalin prevented OA progression by inhibiting pyroptosis in DMM-induced rats. CONCLUSION: These findings indicated that baicalin alleviated chondrocyte injury by upregulating TRIM24 to inactivate the NLRP3/caspase-1-mediated pyroptosis pathway, elucidating the molecular mechanism of baicalin against OA progression.