SIRT1 Protects Against Oxidative Stress-Induced Endothelial Progenitor Cells Apoptosis by Inhibiting FOXO3a via FOXO3a Ubiquitination and Degradation

• Published in: Journal of cellular physiology 2015-09, Vol.230 (9), p.2098-2107
• Main Authors: Wang, Yu-Qiang, Cao, Qing, Wang, Fei, Huang, Li-Ya, Sang, Tian-Tian, Liu, Fang, Chen, Shu-Yan
• Format: Article
• Language: English
• Subjects: Acetylation; Apoptosis; Apoptosis - drug effects; BIM protein; Cell surface; Cells (biology); Centrifugation; Cord blood; Cytology; Deacetylation; Degradation; Dosage; Endothelial Progenitor Cells - drug effects; Fetal Blood; Forkhead Box Protein O3; Forkhead protein; Forkhead Transcription Factors - antagonists & inhibitors; Forkhead Transcription Factors - genetics; FOXO3 protein; Humans; Hydrogen peroxide; Hydrogen Peroxide - administration & dosage; Immunofluorescence; Immunoprecipitation; Low density lipoprotein; Nuclei; Oxidative stress; Oxidative Stress - genetics; Proteins; Proteolysis - drug effects; Signal Transduction - drug effects; SIRT1 protein; Sirtuin 1 - genetics; Stem cells; Substrates; Surface markers; Transcriptional Activation - drug effects; Ubiquitination; Ubiquitination - genetics; Umbilical cord
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.24938

Cell loss due to apoptosis induced by oxidative stress is a major hurdle for endothelial progenitor cells (EPCs)‐based therapy. Sirtuin 1 (SIRT1) plays important roles in many pathophysiological processes by deacetylating various substrates, including forkhead transcription factor (FOXO). However, after deacetylation, the fate of FOXO protein remains to be explored. In the present study, we investigated whether SIRT1 exerted a protective effect on hydrogen peroxide (H2O2)‐induced EPCs apoptosis and, if so, what the underlying mechanism might be. EPCs were isolated and obtained from human umbilical cord blood by density gradient centrifugation and identified by morphology, tube formation ability, cell surface markers, and the ability to take up acetylated low‐density lipoprotein (Dil‐Ac‐LDL) and bind ulex europaeus agglutinin 1 (FITC‐UEA‐1). Immunofluorescence showed that SIRT1 is localized in the nucleus of EPCs in the presence or absence of H2O2. SIRT1 protein level in EPCs was increased by the treatment with H2O2 for 24 h. Incubation of EPCs with H2O2 dose dependently induced EPCs apoptosis. SIRT1 overexpression reduced the rate of EPCs apoptosis induced by H2O2, whereas SIRT1 downregulation and EX527, a specific SIRT1 inhibitor, exerted the opposite effect. SIRT1 overexpression decreased the total FOXO3a protein expression, whereas SIRT1 downregulation and EX527 increased the amount of FOXO3a protein. SIRT1 reduced FOXO3a transcriptional activity according to Bim expression. Co‐immunoprecipitation assay showed that SIRT1 could bind to FOXO3a, reduce its acetylation level and increase its ubiquitination level. To sum up, our work demonstrated that SIRT1 had a pivotally protective role in the regulation of EPCs apoptosis induced by H2O2 and that SIRT1 protected against apoptosis by inhibiting FOXO3a via FOXO3a ubiquitination and subsequent degradation. J. Cell. Physiol. 230: 2098–2107, 2015. © 2015 Wiley Periodicals, Inc.