Objective: To explore the therapeutic effects of Maohuoside A (MHA) on rats with osteoarthritis (OA) induced by anterior cruciate ligament transection, and to analyze its impact on the activation of adenosine monophosphate-activated protein kinase (AMPK). Methods: Sprague-Dawley male rats were divided into six groups: Sham, OA, L-MHA, M-MHA, H-MHA, and H-MHA + Compound C (n = 12 per group). The Sham group had sham surgery, while the OA, L-MHA, M-MHA, and H-MHA groups underwent OA via ACL severing. L-MHA, M-MHA, and H-MHA groups received low, moderate, and high doses of Maohuoside A, respectively, and the H-MHA + Compound C group received high-dose Maohuoside A plus Compound C. Hematoxylin-eosin (HE) staining and Safranin O/Fast Green staining were used to assess articular cartilage damage and degeneration, and the OARSI score was used to evaluate the extent of cartilage degeneration. Results: Compared with the OA group, the L-MHA, M-MHA, and H-MHA groups showed decreased OARSI scores, increased relative levels of Bcl-2 mRNA, decreased relative levels of Bax mRNA, reduced serum levels of IL-1β, IL-6, and TNF-α, increased relative levels of Collagen II mRNA, decreased relative levels of MMP-13 mRNA, and increased relative levels of BMP2, Runx2, and Osterix mRNA. Additionally, the relative level of cartilage AMPKα (Thr172) phosphorylation increased (P<0.05). Conclusion: This study suggests that Maohuoside A exerts anti-osteoarthritis (OA) effects by activating AMPK.