Advanced Glycation End Products Promote PGE2 Production in Ca9-22 Cells via RAGE/TLR4-Mediated PKC–NF-κB Pathway

Advanced glycation end products (AGEs) are compounds that accumulate in hyperglycemic states, contributing significantly to the development of diabetes and its complications, including the exacerbation of periodontal disease. We hypothesized that AGEs affect the expression of inflammatory mediators in gingival cells, thus contributing to the increased severity of periodontitis observed in diabetic patients. Thus, we stimulated the gingival epithelial carcinoma-derived cell line, Ca9-22, with AGEs and examined their effect on the expression of prostaglandin E₂ (PGE₂) and its primary synthesizing enzyme, cyclooxygenase 2 (COX2), key inflammatory mediators in periodontitis. AGEs significantly increased the expression levels of COX2 (n = 6, <i>p</i> < 0.001) and the production of PGE₂ (n = 5, <i>p</i> < 0.05) compared to untreated control and bovine serum albumin (BSA) groups. The receptor for AGEs (RAGE) inhibitor FPS-ZM1 blocked the AGEs-stimulatory effects on COX2 (n = 7, <i>p</i> < 0.01), PGE₂ (n = 6, <i>p</i> < 0.001), and Toll-like receptor 4 (TLR4) expression (n = 7, <i>p</i> < 0.001). Furthermore, AGEs induced the phosphorylation of protein kinase C (p-PKC) via the TLR4 pathway (n = 7, <i>p</i> < 0.01). Crucially, AGEs enhanced NF-κB nuclear accumulation, which was inhibited by blocking either RAGE (n = 5, <i>p</i> < 0.0001) or TLR4 (n = 5, <i>p</i> < 0.0001). In conclusion, these findings demonstrate that AGEs increase PGE₂ production in Ca9-22 cells primarily through a signaling cascade involving RAGE and the TLR4-PKC-NF-κB pathway. Our results suggest TLR4 as a critical mediator that contributes to AGEs-induced inflammation.