GLP-1 inhibits PKCβ2 phosphorylation to improve the osteogenic differentiation potential of hPDLSCs in the AGE microenvironment

Advanced glycation end products (AGEs) have been hypothesized as the etiologic factors of diabetic periodontitis. The discovery of incretins (including GLP-1 and GIP) provides a novel therapy for the treatment of diabetes. Recent reports have shown that glucagon-like peptide-1 (GLP-1) is an important modulator of bone growth and remodeling. The aim of this study was to clarify the mechanism of how GLP-1 weakens/inhibits the effect of AGEs in hPDLSCs (human periodontal ligament stem cells). The hPDLSCs were cultured under simulated conditions of osteogenic culture, AGEs, AGEs + GLP-1, AGEs + GLP-1 + PMA and AGEs + GLP-1 + LY333531. The phenomenon and related mechanism of cell osteogenesis under different microenvironments were evaluated by Alizarin red staining, ALP staining and quantitative activity measurement, RT-qPCR, western blotting and immunofluorescence staining. RT-qPCR showed that AGEs negatively regulated the expression of osteogenic differentiation markers (ALP, BSP, OPN, and Runx2); in contrast, GLP-1 increased the expression of these markers. Furthermore, the expression of RAGE and pPKCβ (PKC phosphorylation) in the AGE group was upregulated, while the expression of RAGE and pPKCβ was decreased in the GLP-1 group compared with the AGE group. AGEs impaired the osteogenic potential of hPDLSCs via PKCβ2. Our phenomenon showed that GLP-1 could reverse the function of AGEs on osteogenic potential. In addition, the mechanism of GLP-1 weakens/inhibits the effect of AGEs in hPDLSCs, possibly by inhibiting PKCβ2 phosphorylation.