This study analysed the effects of different doses of glucosamine hydrochloride (GS-HCl) on cartilage tissue and the levels of joint injury markers in knee osteoarthritis (KOA). The Sham group, KOA group, low-dose GS-HCl group and high-dose GS-HCl group were established, with six mice in each group. The levels of joint injury markers (COMP, CS846 and CTX-II), inflammatory cytokines (IL-6, TNF-α and iNOS), oxidative stress indicators (MDA and SOD) and matrix remodelling proteins (MMP-3 and TIMP-1) were analysed. The degeneration of knee femoral condyles, histopathological changes and tissue apoptosis rate of the articular cartilage was also assessed. Mice in the KOA group displayed elevated COMP, CS846, CTX-II, IL-6, TNF-α, iNOS and MDA contents, reduced SOD activity, an irregular articular cartilage surface, a serious cartilage defect, a disordered articular cartilage surface in the defect, disappeared cartilage cells, obvious synovial cell proliferation and visible inflammatory cell infiltration. In the tissue, apoptosis rate and MMP-3 and TIMP-1 protein expression increased. Different doses of GS-HCl treatment could reduce COMP, CS846, CTX-II, IL-6, TNF-α, iNOS and MDA contents, apoptosis rate and MMP-3 and TIMP-1 protein expression, increase SOD activity and improve histopathological conditions in KOA mice. The improvement effects in each indicator in the high dose-GS-HCl group were more significant than those in the low dose-GS-HCl group. The intragastric administration of the GS-HCl group partially prevents the degeneration of articular cartilage in KOA mice. The mechanism may be to reduce inflammatory factors and oxidative stress indicator expression and matrix degradation, thereby delaying osteoarthritis progression.