The Effects Of Liraglutide On Human Periodontal Ligament Cells Induced By Advanced Glycosylation End Products

Objective: Diabetes as a systemic promote factors become a high risk factor of periodontal disease in recent years.Advanced glycosylation end products(AGEs)have been proved to be a central role in diabetes complications.A large number of studies have demonstrate that AGEs could promote the periodontal ligament cells apoptosis,suppress the osteogenetic differentiation and promote the formation of inflammation factor,resulting in destruction of periodontal tissue.Liraglutide(Lira)is a novel diabetes drugs.In this study,we aim to detect the function of Lira on periodontal ligament cells induced by AGEs through apoptosis,osteogenesis and inflammatory factor expression.Discussing the therapeutic effects of Lira on periodontal disease with diabetes.Methods: Primary cultures were obtained by enzyme digestion method combined tissue block cultivation method.Selecting passage 3 to passage 6 periodontal ligament cells(PDLCs)in the following experiments.Immunohistochemistry and Alizarin red staining were used to identify the PDLCs.Method MTT technique was used to observe the optimal concentration and action time of AGEs.Optimal concentration and action time of Lira to the PDLCs induced by AGEs were determined with MTT.Method MTT technique was used to test the proliferative activity of Lira to the PDLCs induced by AGEs.Real-time PCR and western blot was used to detect the expression level of RAGE and GLP-1R.Annexin-V-FITC/PI double-staining assay and Hoechst33258 dye staining were used to explore apoptosis.Alizarin red staining,Real-time PCR and Western blot were used to detect the potential of osteogenesis.Real-time PCR and Western blot were used to detect the expression levels of inflammatory markers.Results:(1)Immunocytochemical showed that cells with spindle shape were positive in vimentin and negative in keratin.(2)MTT showed that the optimal concentration of AGEs was 100 μg/m L.The optimal concentration of Lira to the PDLCs induced by AGEs was 100 ng/m L,the action time was 24 h.he proliferation inhibition rate of control group was lower than that of AGEs group,The proliferation inhibition rate of Lira group was lower than the control group,AGEs + Lira group was lower than AGEs group.(3)Western blot and Real-time PCR showed that the expression level of RAGE in AGEs group was higher than that in control group,Lira group was lower than the control group,AGEs + Lira group was lower than AGEs group.The expression level of GLP-1R in Lira group was higher than that in the control group,AGEs group was lower than the control group,AGEs + Lira group was higher than AGEs group.(4)Annexin-V-FITC/PI double-staining assay and Hoechst33258 dye staining showed that the apoptosis rates of AGEs group was higher than that in the control group,Lira group was lower than the control group,AGEs + Lira group was lower than AGEs group.(5))Western blot and Real-time PCR showed that the expression level of ALP and Runx2 and the number of mineralized nodule formation in AGEs group was lower than that in the control group,AGEs group was lower than the control group,AGEs + Lira group was higher than AGEs group.The expression level of IL 6 and TNF – α in AGEs group was lower than that in the control group,AGEs group was lower than the control group,AGEs + Lira group was higher than AGEs group.Conclusion:(1)The optimal concentration of Lira to the PDLCs induced by AGEs was 100 nmol /m L,the action time was 24 h.(2)RAGE and GLP-1R were expressed on PDLCs.(3)Lira suppressed the expression of RAGE of normal PDLCs and PDLCs induced by AGEs.Lira promoted the expression of GLP-1R of normal PDLCs and PDLCs induced by AGEs.(4)Lira promoted the normal PDLCs and PDLCs induced by AGEs proliferation and osteogenic differentiation.It also inhibit the normal PDLCs and PDLCs induced by AGEs apoptosis and down-regulate the expression of proinflammatory.