Osteoarthritis (OA) causes progressive cartilage and bone degradation for which no disease-modifying therapy exists. Withania somnifera (WS) and Yogaraj Guggulu (YG) are centuries-old herbal formulation used in Ayurveda for joint disorders, but its molecular mode of action is poorly defined. We performed label-free LC-MS/MS proteomics on rat knee-joint tissue to uncover pathways modulated by oral WS and YG in an MIA-induced OA model. Eighty male Sprague-Dawley rats were randomised into normal control, vehicle, disease control, diclofenac (standard), three YG dose groups (low, medium, high) and three WS groups (low, medium, high). WS or YG or vehicle was given orally for 21 days after OA induction (5 mg MIA, intra-articular). Knee joints were harvested on Day 28, flash-frozen, pulverised and processed for LC-MS/MS. Each biological replicate was injected with three times, resulting in 12 replicates per group. WS and YG normalised multiple OA-dysregulated proteins, dampening NF-κB/IL-17 signalling and collagen-degradation cascades while restoring metabolic and cytoskeletal processes, congruent with micro-CT, histology and behavioural recovery.