| Objective To explore whether intracellular reactive oxygen species (ROS) participating in senescence of bone marrow mesenchymal stem cells (BMSCs) from patients with systemic lupus erythematosus (SLE) and invstgate the mechanism that controls the intracellular ROS levels.Methods Human bone marrow aspirates were collected from iliac of ten donors and ten SLE patients and cultured in vitro. Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was tested by fluorescence microscope. BMSCs were monitored using the Senescence Associated-galactosidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expressions of FoxO3, phospho-FoxO3 (p-FoxO3), AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis.Results There were no differences in morphology and nuclear size of BMSCs between SLE patients and normal controls. The percentage of SAβ-gal positive BMSCs from SLE patients was higher than that from normal controls (p<0.05) Intracellular ROS levels of BMSCs from SLE patients increased more significantly than that of normal controls in vitro (p<0.05). No significant differences in the expressions of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a increasing trendency. The expressions of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels (p<0.05).Conclusion:These data suggested that BMSCs from SLE patients aged more quickly, with high SAβ-gal activity and up-production of intracellular ROS, accompanied by up-regulation of p-FoxO3 and p-AKT at protein levels, which might have a tight link with the pathogenesis of SLE by maintaining BMSCs lifespan.
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