Dysfunction Of Human Adipose Tissue-derived Stem Cells Induced By Advanced Glycation End Products And The Involved Mechanisms

Objective: To study the effects of advanced glycation end products-human serum albumin (AGE-HSA) on the oxidative stress, proliferation, migration and serection of adipose tissue derived stem cells(ADSCs), and the involved molecular mechanisms.Methods: The subcutaneous adipose tissue samples were obtained from female patients receiving cosmetic surgery, then enzymatically digested and centrifugated in vitro. The ADSCs were determined by fluorescence activated cell sorter (FACS) analysis and in vitro differentiation induced according to different previous strategies. After stimulated with different dosages of AGE-HSA, the expression of RAGE, MAPK and NF-κB was determined by western blot. Then the cells were preincubated with ERK and p38 inhibitors before the stimuli, the expression of RAGE, Rev-erbα, Bmal1 and Per2 mRNA at the different timepoints were determined by Real-time PCR, FCM and CCK8 were used to evaluate the proliferation capability, oxyblot assay and FSM were used to investigate the oxidative stress, transwell and in vitro scratch assay were used to determine the migration capability, and secreted cytokines were measured by ELISA. The role of Rev-erbα, a key molecular of rhythm genes system, in the pathological effects of AGE-HSA was investigated by transfection with siRNA aiming at such gene before the stimulus with AGE-HSA.Results: AGE-HSA could significantly inhibit the migration of ADSCs, but promote the proliferation and secretion of such cells, which accompalished the increased oxidative stress and activation of RAGE-MAPK-NF-κB pathway. ERK inhibitor, p38 inhibitor and rosuvastatin could amerolate the pathological effects of AGE-HSA to different extents. AGE-HSA could also induce the dyssynchronization of rthym system including Rev-erbα, Bmal1 and Per2. Inhibitory effects on the migration of such cells could be attenuated by transfection with Rev-erbαsiRNA. Conclusion: AGE-HSA could induce dysfunction of ADSCs, which might be mediated by activation of RAGE-MAPK-NF-κB pathway, increased oxidative stress and expression of key molecular Rev-erbα, then the dyssynchronization of rthym genes. Promoted proliferation and secretion of ADSCs after the acute exposure to AGE-HSA might be the self protection mechanism of such cells against the outside negative stimuli. These findings partly revealed a novel mechanism about rhythm gene Rev-erbαparticipating in the dysfunction of ADSCs induced by AGE-HSA.