Spermidine Retarded the Senescence of Multipotent Mesenchymal Stromal Cells In Vitro and In Vivo through SIRT3-Mediated Antioxidation

Figure 7

The long-term high-spermidine diet hindered rADMSC senescence in vivo. (a) A representative micrograph showing morphological changes of primary rADMSCs from different groups at the indicated time points in vitro after a one-year high-spermidine diet;  μm. (b) A representative image showing intracellular ROS levels by molecular probe H2DCFDA staining in hUCMSCs with indicated treatment;  μm. (c, d) Quantification of intracellular ROS levels was performed by fluorescence-activated cell sorting (FACS) analysis of stained cells to obtain the mean fluorescence intensity. (e) A representative image showing SA-β-gal staining in rADMSCs at PDL4, PDL8, and PDL16 with indicated groups, respectively,  μm. (f) The percentage of senescent cells stained in blue in each group was assessed by quantitative analysis. (g) Representative western blotting results showing the protein expression levels of GLR1, SIRT3, OCR4, SOX2, p-P53-ser15, P53, or P21 in different groups. (h) Expression levels of the corresponding proteins were quantified using ImageJ software and normalized to the control group. Approximately 10⁵ cells per well were seeded in 6-well plates. All quantitative data were obtained from three independent experiments and presented as ; and represent a significant difference between the indicated groups.

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