Modified Qianghuo Shengshi Decoction Ameliorates Osteoarthritis via Inhibiting <scp>PI3K</scp>/Akt Pathway‐Related Ferroptosis

Modified Qianghuo Shengshi decoction (MQSD), a TCM formula, is clinically used for osteoarthritis (OA) symptom relief. Its exact molecular actions, however, are not fully understood. This investigation seeks to elucidate the molecular mechanisms by which MQSD impedes OA progression. High performance liquid chromatography (HPLC) was employed to delineate MQSD's chemical profile. The CCK-8 assay determined MQSD's impact on chondrocyte viability in the presence or absence of IL-1β. EDU and Annexin V-FITC assays evaluated chondrocyte proliferation and apoptosis, respectively. Alcian Blue staining probed extracellular matrix (ECM) secretion by chondrocytes. Network pharmacology was utilised to pinpoint disease targets, while various assays assessed ferroptosis-related chondrocyte phenotypes under Erastin or Ferrostatin-1 treatments. IHC, qRT-PCR and western blot analyses were conducted to ascertain MQSD's cellular effects. Micro-CT and Safranin O Fast Green staining provided insights into knee joint morphology and cartilage integrity. Molecular docking assessed the binding affinity of MQSD's active compounds to PI3K/Akt/GPX4. MQSD promotes chondrocyte proliferation, prevents apoptosis, and enhances viability. It further modulates the ECM output and the anabolic-to-catabolic ratio within chondrocytes. Through network pharmacology, ferroptosis linked to the PI3K/Akt pathway is identified as crucial. Pre-application of MQSD elevates the GSH/GSSG ratio and mitochondrial density, while it reduces Fe²⁺ levels, ROS, and lipid droplet accumulation in chondrocytes. Moreover, MQSD activates GPX4 and inhibits p-AKT levels. Molecular docking studies affirm the strong interaction between MQSD's key ingredients and the PI3K/Akt/GPX4 pathway components. Through modulation of ferroptosis via the PI3K/Akt pathway, MQSD retards OA progression.