C1GALT1 plays a pivotal role in O-glycan modification. We observe an increase in both the mRNA and protein levels of C1GALT1 in ATDC5 cells stimulated with IL-1β and TGF-β, which are the major inflammatory factors in osteoarthritis. This trend is also mirrored in the cartilage tissue of mice with experimentally induced arthritis. When we suppress the mRNA of C1GALT1, we observe a decrease in the mRNA expression levels of the inflammatory genes, nitric oxide synthase (iNOS), and type II collagen (Collagen II). This result indicates that C1GALT1 is part of a regulatory pathway that promotes inflammation. Our analysis on the NetOGlyc-4.0 platform, which leverages extensive data, reveals potential O-glycosylation sites in both mouse and human IL-1β receptors. It suggests that C1GALT1 mediated O-glycosylation on IL-1 receptors would lead to an increase in iNOS expression, thereby fostering inflammation. Inhibition of C1GALT1 mRNA results in a reduction of IL-1R1 protein expression. Additionally, when we inhibit either the mRNA or protein of C1GALT1, we observe a decrease in the expression of IL-1R1 in the perinuclear endomembrane system. This underscores the significance of C1GALT1-mediated glycosylation in regulating the formation of IL-1R1, and inhibiting C1GALT1 may potentially alleviate arthritis in vivo. Notably, Itraconazole, which inhibits C1GALT1, exerts minimal harm to the human body. In light of this, Itraconazole emerges as a prospective candidate for arthritis treatment in innovative applications.