Osteoarthritis (OA) is a chronic joint disease characterized by the deterioration of cartilage and subchondral bone in the joints. Currently, there is no complete cure for OA, only treatments designed to temporarily relieve pain and improve function. Compared with the high cost of surgical treatment, medical treatment of OA is more acceptable and cost-effective. Rutin, as a flavonoid, has been shown to have anti-OA properties. We evaluated the effects of rutin on chondrocytes in lipopolysaccharide (LPS)-induced OA and on OA in rats induced by anterior cruciate ligament transection. We found that rutin effectively reduced the expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and matrix metalloproteinase 13 (MMP-13) and increased the expression of Col II and aggrecan ( < 0.001). In addition, we also found that rutin increased the expression of cystathionine-β-synthase (CBS) and inhibited the expression of Rho-related coiled-coil protein kinase (ROCK) in chondrocytes ( < 0.05), thereby effectively inhibiting the inflammatory progression of OA. We concluded that rutin inhibits the inflammatory progression of OA through the -mediated signaling pathway.