MON-107 Endothelium Derived Factors Influence Differentiation of Fat Derived Stromal Cells

Abstract

By 2030, the prevalence of diabetes will rise by 54% and will become a global epidemic. Aerobic exercise is known to show positive effects on cardiovascular health in diabetes and pre-diabetes subjects. However, the effect of exercise on vascular health was being studied using plasma biochemistry rather than cellular data. We investigated if endothelial conditioned media can affect MSC differentiation in adipogenic environment. We also examined the effect of aerobic exercise on the subject’s fat derived MSCs to study stromal cell differentiation. Methods: In vitro, commercially obtained human fat-derived stromal cells were exposed to adipogenic media and endothelial conditioned media (obtained from mature endothelial cells, HUVECs, 40 fold concentrated) was added. MSC differentiation was monitored by RT-PCR. For the in-vivo study, five prediabetic subjects, BMI > 25 were enrolled in a 12-week exercise program. Parameters were assessed pre and post exercise. Stromal cells were obtained from subcutaneous abdominal fat and cultured for 2-3 weeks. Stromal cells were then analyzed for gene expression. Results: We noted a reduced expression of markers for adipogenic differentiation such as PL1N, FABP4, CEBPA and PPARG (24.6, 13.3, 9.8, 4.2 fold, p= 0.00001, 0.02, 0.00001, 0.0001 respectively). We also observed increased expression of bone formation markers RUNX2 and BMP2 (1.5, 1.3 fold, p=0.01, 0.01 respectively) which were increased significantly. We also noted upregulation of VEGF and KDR (1.6 and 2.3 fold, respectively). When we analyzed mRNA expression of patient’s fat-derived stromal cells, we noticed, an upregulation of glucose transporter GLUT1, GLUT4 (3.6, 1.14 fold). Gene expression of most of the antioxidants such as SOD2, SOD3, GPX1, GPX3 and CAT (1.5, 1.7. 1.16, 1.6 and 1.6 fold respectively) increased post exercise with a statistically significant upregulation of SOD1, a cytosolic antioxidant (p= 0.04, 1.8 fold). Differentiation towards fat, bone, muscle and cartilage, using mRNA expression analysis, showed a trend of upregulation of bone formation marker osteocalcin (BGLAP), BMP6, SMAD3 and TGF-ᵝ (1.3, 2.6, 2, 3.1 fold respectively) post exercise. Concomitantly, serum values of osteocalcin increased significantly, post-exercise. Biophysical and biochemical parameters demonstrated that exercise increased METs significantly (p=0.04, 12%) while reduced body weight, systolic, diastolic blood pressure, HbA1c, cholesterol by 4.2%,9%, 9%, 10%, 9.8 % respectively (p=0.04, 0.04, 0.05, 0.001, 0.0003 respectively) Conclusion: Both in-vivo and in-vitro data showed that exercise augments cellular glucose transporters, anti-oxidants and reduce MSC inflammation. Addition of endothelium-derived factors and exercise both appear to augment bone formation markers indicating a cross-talk between endothelium, fat differentiation, and bone formation, post-exercise.