RAGE and TGF-β1 Cross-Talk Regulate Extracellular Matrix Turnover and Cytokine Synthesis in AGEs Exposed Fibroblast Cells

AGEs accumulation in the skin affects extracellularmatrix (ECM) turnover and triggers diabetes associated skin conditions and accelerated skin aging. The receptor of AGEs (RAGE) has an essential contribution to cellular dysfunction driven by chronic inflammatory responses while TGF-beta 1 is critical in both dermal homeostasis and inflammation. We investigated the contribution of RAGE and TGF-beta 1 to the modulation of inflammatory response and ECM turnover in AGEs milieu, using a normal fibroblast cell line. RAGE, TGF-beta 1, collagen I and III gene and protein expression were upregulated after exposure to AGEs-BSA, and MMP-2 was activated. AGEs-RAGE was pivotal in NF-kappa B dependent collagen I expression and joined with TGF-beta 1 to stimulate collagen III expression, probably via ERK1/2 signaling. AGEs-RAGE axis induced upregulation of TGF-beta 1, TNF-alpha and IL-8 cytokines. TNF-alpha and IL-8 were subjected to TGF-beta 1 negative regulation. RAGE's proinflammatory signaling also antagonized AGEs-TGF-beta 1 induced fibroblast contraction, suggesting the existence of an inhibitory cross-talk mechanism between TGF-beta 1 and RAGE signaling. RAGE and TGF-beta 1 stimulated anti-inflammatory cytokines IL-2 and IL-4 expression. GM-CSF and IL-6 expression appeared to be dependent only on TGF-beta 1 signaling. Our data also indicated that IFN-gamma upregulated in AGEs-BSA milieu in a RAGE and TGF-beta 1 independent mechanism. Our findings raise the possibility that RAGE and TGF-beta 1 are both involved in fibrosis development in a complex cross-talk mechanism, while also acting on their own individual targets. This study contributes to the understanding of impaired wound healing associated with diabetes complications.