IADR Abstract Archives
MiR-375-3p Promotes Macrophage M1 Polarization and Represses BMSCs Osteogenic Differention by Suppressign PDK1 in Periodontic Tissue of Diabetic Rats
Objectives: This study aims to investigate the role of miR-375-3p in the progression of diabetic periodontitis and elucidate the underlying mechanism.
Methods: The diabetic rat model is constructed by STZ intraperitoneal injection and periodontitis model is built by silk ligament and lipopolysaccharide (LPS) injection. MiR-375-3p level in plasma of diabetic rats is analyzed by qRT-PCR. Single molecule localization microscopy is applied to track miR-375-3p in plasma and periodontic tissue. The target gene is found by bioinformatic tools and verified by dual-luciferase reporter gene testing. DCFH-DA staining to evaluate the content of reactive oxygen species (ROS). Verifying the M1 polarization of macrophages by flow cytometry. ALP activity test and Alizarin staining are utilized to evaluate the osteogenic differentiation of Bone Marrow Stromal Cell (BMSCs). Western blotting and qRT-PCR are used to test the transcriptional and expressional level miR-375-3p and its target genes.
Results: The results demonstrate that miR-375-3p is upregulated in the plasma of diabetic rat and miR-375-3p is uptaken by BMSCs and macrophages in periodontic tissue. Database searching verify Pdk1 as the target gene of miR-375-3p. MiR-375-3p inhibits the expression of PDK1 and downregulate the PDK1-AKT signal path, results into the accumulation of ROS, promotes the M1 polarization of macrophages and repress the osteogenic differentiation of BMSCs.
Conclusions: The upregulated miR-375-3p in plasma of diabetic rats may exacerbate the progress of periodontitis by targeting Pdk1 and downregulating PDK1-AKT signal path, our study offers a potential therapeutic target of diabetic periodontitis.
Methods: The diabetic rat model is constructed by STZ intraperitoneal injection and periodontitis model is built by silk ligament and lipopolysaccharide (LPS) injection. MiR-375-3p level in plasma of diabetic rats is analyzed by qRT-PCR. Single molecule localization microscopy is applied to track miR-375-3p in plasma and periodontic tissue. The target gene is found by bioinformatic tools and verified by dual-luciferase reporter gene testing. DCFH-DA staining to evaluate the content of reactive oxygen species (ROS). Verifying the M1 polarization of macrophages by flow cytometry. ALP activity test and Alizarin staining are utilized to evaluate the osteogenic differentiation of Bone Marrow Stromal Cell (BMSCs). Western blotting and qRT-PCR are used to test the transcriptional and expressional level miR-375-3p and its target genes.
Results: The results demonstrate that miR-375-3p is upregulated in the plasma of diabetic rat and miR-375-3p is uptaken by BMSCs and macrophages in periodontic tissue. Database searching verify Pdk1 as the target gene of miR-375-3p. MiR-375-3p inhibits the expression of PDK1 and downregulate the PDK1-AKT signal path, results into the accumulation of ROS, promotes the M1 polarization of macrophages and repress the osteogenic differentiation of BMSCs.
Conclusions: The upregulated miR-375-3p in plasma of diabetic rats may exacerbate the progress of periodontitis by targeting Pdk1 and downregulating PDK1-AKT signal path, our study offers a potential therapeutic target of diabetic periodontitis.