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BACKGROUND: Acupotomy is an effective method for the clinical treatment of osteoarthritis, with affirmed clinical outcomes, but the specific mechanisms remain unclear OBJECTIVE: To investigate the role of acupotomy in modulating chondrocyte autophagy to promote chondrocyte homeostasis in osteoarthritis. METHODS: Twenty-eight New Zealand rabbits were randomly divided into control group, osteoarthritis group, acupotomy group, and hyaluronic acid group, with seven rabbits in each group. The knee osteoarthritis rabbit model was prepared using the Videman method in the latter three groups. After modeling, the control group and osteoarthritis group received no interventions. The acupotomy group received acupotomy treatment 15 minutes per time, once a week, while the hyaluronic acid group received intra-articular injection of hyaluronic acid once a week, with a continuous treatment duration of 5 weeks. The day after the final intervention, knee joint macrostructure was observed using DR imaging, chondrocyte ultrastructure was examined through transmission electron microscopy, apoptosis of chondrocytes was assessed using Tunel staining, and western blot analysis was used to detect the expression of proteins related to the PI3K/Akt/mTOR pathway. RESULTS AND CONCLUSION: The DR imaging results revealed that the osteoarthritis group exhibited narrowed knee joint spaces and the formation of periarticular osteophytes, while the hyaluronic acid group and acupotomy group showed widened knee joint spaces with a reduction in periarticular osteophytes. Transmission electron microscopy results demonstrated a decreased number of autophagosomes in chondrocytes in the osteoarthritis group, along with nuclear shrinkage, nuclear membrane rupture, incomplete organelle morphology, and a clear tendency towards cell death. In contrast, both the hyaluronic acid group and acupotomy group exhibited a significant increase in autophagosomes, intact nuclear membranes, and a well-preserved cellular state. Tunel staining results indicated a considerable decrease in the number of apoptotic cells in the hyaluronic acid group and acupotomy group compared with the osteoarthritis group. Western blot results revealed that, compared with the control group, the expression levels of Beclin1, Cath D, and LC3II/LC3I were significantly decreased in the osteoarthritis group (P < 0.05), while the expression levels of p-Akt/Akt and p-mTOR/mTOR were significantly increased ( P < 0.05); compared with the osteoarthritis group, the expression levels of Beclin1, Cath D, and LC3II/LC3I were significantly increased in both the hyaluronic acid group and acupotomy group (P < 0.05), while the expression levels of p-Akt/Akt and p-mTOR/mTOR were significantly decreased (P < 0.05). To conclude, acupotomy intervention can modulate the PI3K/Akt/mTOR signaling pathway to enhance the autophagic level in chondrocytes, thereby maintaining chondrocyte homeostasis. This ultimately leads to a slowdown in cartilage degeneration. [ABSTRACT FROM AUTHOR]