The medicine and food homologous plant Ligustri Lucidi fructus attenuates joint capsule fibrosis in post-traumatic joint contracture through machine learning guided transcriptomic and in vivo validations

Purpose Post-traumatic joint contracture (PTJC) is a disabling complication characterized by joint capsule fibrosis. This study investigated the anti-fibrotic effects and mechanisms of Ligustri Lucidi Fructus (LLF), a medicine and food homologous plant, against PTJC using integrated in vivo experiments, network pharmacology, and machine learning. Methods A rat PTJC model was established to evaluate LLF efficacy via range of motion (ROM) measurements and histological staining. Active ingredients and targets of LLF were identified using public databases, followed by protein-protein interaction network construction and functional enrichment analyses. LASSO regression and Random Forest algorithms were applied to an independent RNA-sequencing dataset to pinpoint core genes, which were further evaluated by molecular docking. Predicted signaling pathways and myofibroblast activation were verified using immunofluorescence and Western blotting. Results LLF dose-dependently improved joint ROM, reduced swelling, and alleviated collagen deposition and myofibroblast activation. Network pharmacology identified 9 active compounds and 113 overlapping targets. Machine learning and RNA-seq validation pinpointed five core genes driving PTJC: Tnf , Nfkb1 , Akt1 , Tgfb1 , and Ptgs2 . Molecular docking confirmed strong binding affinities between key compounds and these targets. Experimental validation revealed that LLF significantly inhibited the TNF-α/NF-κB, PI3K/Akt, and TGF-β1/Smad pathways. Conclusion LLF ameliorates PTJC by suppressing inflammation and fibroblast activation through multi-target, multi-pathway synergy that interrupts the inflammation-fibrosis vicious cycle, primarily via the TNF-α/NF-κB, PI3K/Akt, and TGF-β1/Smad pathways. These findings support the clinical potential of this medicine and food homologous plant against fibrotic joint diseases.