Bone marrow mesenchymal stem cell-derived exosomes reverse high glucose-induced osteogenic dysfunction of bone marrow mesenchymal stem cells via PI3K/AKT signaling pathway
TAO Endong,XIA Weijie, WANG Fulin, WANG Xianyu, CAI Leyi, FENG Yongzeng.
Department of Orthopedic Surgery, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, China
TAO Endong,XIA Weijie,WANG Fulin, et al. Bone marrow mesenchymal stem cell-derived exosomes reverse high glucose-induced osteogenic dysfunction of bone marrow mesenchymal stem cells via PI3K/AKT signaling pathway[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2024, 54(8): 641-649.
Abstract：Objective: To investigate the therapeutic effect of bone marrow mesenchymal stem cell-derived exosomes (BMSCs-EXOs) on high glucose-induced osteogenic dysfunction of bone marrow mesenchymal stem cells (BMSCs) and its mechanism. Metheds: Exosomes were extracted from BMSCs by ultracentrifugation, and
their characteristics were evaluated by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot. High glucose medium was used to simulate the high glucose microenvironment, and the effect of BMSCs-EXOs on the oxidative stress level, osteogenic differentiation, and mineralization ability of high glucose-induced BMSCs were studied by DCFH-DA fluorescent probe, Western blot, cell immunofluorescence (IF), alkaline phosphatase staining (ALP), and alizarin red staining (ARS). In addition, the PI3K pathway blocker (LY294002) was used to inhibit the phosphorylation of PI3K to study whether BMSCs-EXOs affect the osteogenic differentiation and mineralization ability of BMSCs by activating the PI3K/AKT signaling pathway. Results:The successfully extracted exosomes from BMSCs were evaluated by TEM, NTA and Western blot. Compared with the control group, the oxidative stress level was significantly increased in the HG group (P<0.05) and the protein expression levels of HO-1 and NQO-1 significantly decreased (P<0.05); the activities of ALP and ARS were significantly decreased (P<0.05), and the protein expression levels of COL1A1, RUNX2, BMP2, p-PI3K and p-AKT were significantly decreased (P<0.05). Compared with the HG group, the oxidative stress level was
significantly decreased (P<0.05) and the protein expression levels of HO-1 and NQO-1 were significantly upregulated (P<0.05); the activities of ALP and ARS were significantly increased (P<0.05), and the protein expression levels of COL1A1, RUNX2, BMP2, p-PI3K and p-AKT were significantly up-regulated (P<0.05).When LY294002 was added, the phosphorylation expression of the PI3K/AKT signaling pathway was significantly inhibited, and the osteogenic effect of BMSCs-EXOs was also significantly weakened (P<0.05). Conclusion:BMSCs-EXOs partially restored osteogenic differentiation and mineralization by reducing high glucose-induced oxidative stress levels, and this osteogenic effect of BMSCs-EXOs was demonstrated to be achieved through the PI3K/AKT signaling pathway.
