Mechanism Of Ascorbic Acid In Regulating The Senescence Of Bone Marrow Mesenchymal Stem Cells

Objective:To study the specific role of ascorbic acid in delaying BMSCs senescence and its potential molecular mechanism by constructing the Bone Marrow Mesenchymal Stem Cell(BMSCs)aging model in vitro.Methods:1.The Seven-day-old SD rats were selected and the bone marrow of the tibia and fibula was extracted under aseptic conditions.The whole bone marrow adherent culture was used to isolate and culture BMSCs in vitro.2.BMSCs were induced to co-culture with D-gal to induce the aging of BMSCs,and to establish the aging model of BMSCs in vitro.3.The ascorbic acid was co-cultured with the senescent BMSCs.The changes of the positive rate of SA-?-Gal staining after treatment of ascorbic acid were detected by SA-?-Gal staining.The intracellular changes of ROS content were detected by immunofluorescence microscope after treatment of ascorbic acid.The changes of aging-related protein p16INK4a and alterations of related signal pathways were detected by Western blot analysis after treatment of ascorbic acid.4.Combined with AKT inhibitor and ascorbic acid to further verify the AKT/mTOR signaling pathway in the process of the role of ascorbic acid in restraining the aging of BMSCs.Results:D-galactose can induce the premature aging of BMSCs,meantime,the ROS content of aging BMSCs increased.Ascorbic acid can reduce the generation of senescent cells in aged BMSCs,along with the decreases of level of ROS in senescent BMSCs and the decreases of the content of p16INK4a.The ascorbic acid affects the changes of AKT/mTOR signaling pathway,too.At last the combination of AKT inhibitor can increase the effect of ascorbic acid in restraining the aging of BMSCs.Conclusions:Ascorbic acid can delay the senescence of ascorbic acid by inhibiting intracellular ROS formation and affecting intracellular AKT/mTOR signaling pathway.