Nicotinamide adenine dinucleotide rejuvenates septic bone marrow mesenchymal stem cells

Figure 5 Nicotinamide adenine dinucleotide treatment rescue function of septic bone marrow mesenchymal stem cells. A: Representative images of EdU incorporation, EdU (magenta), and DAPI (blue) staining. The percentage of EdU-positive cells increased after treatment with nicotinamide adenine dinucleotide (NAD) [healthy donors (HD): n = 7; sepsis: n = 6; sepsis with NAD: n = 6]; B: Cellular senescence assessed by β-gal staining in bone marrow mesenchymal stem cells (BMSCs) from three groups; C and D: Alizarin Red S staining (original magnification, × 4) and quantification of calcium precipitation; E: Quantitative real-time polymerase chain reaction analysis of osteogenic genes (runt-related transcription factor 2, alkaline phosphatase, and Sp7 transcription factor 7) in HD BMSCs (n = 5), septic BMSCs (n = 5), and NAD-treated septic BMSCs (n = 5); F and G: Oil Red O staining (original magnification, × 10) and lipid droplet quantification; H: Quantitative real-time polymerase chain reaction analysis of adipogenic genes (peroxisome proliferator-activated receptor gamma, fatty acid binding protein 4, and perilipin-1) in HD BMSCs (n = 5), septic BMSCs (n = 5), and NAD-treated septic BMSCs (n = 5). Results were presented as fold change relative to undifferentiated BMSCs. Error bars represent the mean ± SEM. P values were determined using One-Way ANOVA (^aP < 0.05, ^bP < 0.01, ^cP < 0.001). HD: Healthy donors; NAD: Nicotinamide adenine dinucleotide; RUNX2: Runt-related transcription factor 2; ALP: Alkaline phosphatase; SP7: Sp7 transcription factor 7; PPARG: Peroxisome proliferator-activated receptor gamma; FABP4: Fatty acid binding protein 4; PLN1: Perilipin-1.