Effect of SIRT6/NF-kappa B signal axis during hematopoietic stem and progenitor cell senescence induced by tert-butylhydroperoxide in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND:SIRT6/NF-κB is the important signal axis to cellsenescence, but the effect of SIRT6/NF-κB signal axis to hematopoietic stem and progenitor cell(HSC/HPC) senescence is unclear. OBJECTIVE:To induce (t-BHP) in vitro and to investigate the role of SIRT6/NF-κB signal axis in Sca-1+HSC/HPC senescence induced by tert-butylhydroperoxide in vitro. METHODS:Sca-1+HSC/HPC was isolated and purified by magnetic activated cellsorting. Sca-1+HSC/HPC senescence was induced by 100μmol/L tert-butylhydroperoxide in vitro. The senescence-associatedβ-Galactosidase staining, cellcycle analysis and culture of mixed hematopoietic progenitor cellwere used to investigate the biological effects of tert-butylhydroperoxide on Sca-1+HSC/HPC senescence. The expression of senescence associated SIRT6, NF-κB mRNA and protein was examined by real-time fluorescence quantitative PCR and western blot assay. RESULTS AND CONCLUSION:Compared with control group, the percentage of positive cells expressing SA-β-Gal and cells in G0/G1 phase increased and the number of forming colony of mixed hematopoietic progenitor decreased in the aging group. It showed lower expression of SIRT6 and higher expression of NF-κB in the aging group. The SIRT6/NF-κB signal axis may play a key role in the Sca-1+ HSC /HPC senescence inducted by tert-butylhydroperoxide.