Network Pharmacological Prediction and Experimental Analyses Reveal That Naringin Alleviates Osteoarthritis Progression by Targeting MMP13.

Abstract

Osteoarthritis (OA) is a severe chronic inflammatory disorder with limited treatment options. Naringin (nar) has been shown to protect against OA; however, its mechanisms of action on OA remain poorly understood. This study aims to investigate the molecular mechanism of nar in treating OA via network pharmacology and experiments. Differentially expressed genes (DEGs) were identified using GSE283079 dataset. Protein-protein interaction (PPI) network was constructed using STRING database, and protein interactions were analyzed. Network pharmacology was employed to investigate the molecular interaction network influenced by nar in OA, and molecular docking was applied to predict the binding interactions between nar and core genes. The OA mouse models were constructed using anterior cruciate ligament transection (ACLT) to explore the action of nar in vivo. The OA damage was examined using Hematoxylin and Eosin (HE) and Safranin-O/Fast Green staining, along with Osteoarthritis Research Society International (OARSI) scoring for quantitative histopathological evaluation. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) positive rate and inflammation factor (tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta), and reactive oxygen species (ROS) levels were detected using corresponding assay kits. The protein expression was analyzed using western blot. Cell viability and cell apoptosis were examined using cell counting kit 8 (CCK8) assay kit and flow cytometry assays. In GSE283079 dataset, the up-regulation of DEGs was enriched in immune response activation, cartilage development, collagen metabolic process, and leukocyte proliferation. Additionally, matrix metalloproteinase 13 (MMP13), MMP1, and phospholipase A2 group IIA (PLA2G2A) may be the core genes for nar-protected OA. The binding energy of nar and MMP13 was strongest. In vivo OA models, nar mitigated OA progression and reduced OARSI scores. Mechanistically, nar suppressed cell apoptosis, inflammation factor productions, extracellular matrix (ECM) degradation, and ROS production via decreasing MMP13. Nar alleviates OA malignant progression via reducing MMP13. Key words Osteoarthritis " Naringin " Network pharmacology " MMP13 " Molecular mechanism.