›› 2009, Vol. 29 ›› Issue (10): 1182-.

• Original article (Basic research) • Previous Articles     Next Articles

Oxidative mechanism of homocysteine-induced apoptosis in endothelial progenitor cells

BAO Xiao-mei, WU Chun-fang, LU Guo-ping   

  1. Department of Cardiology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2009-10-25 Published:2009-10-26


Objective To investigate the oxidative mechanism of homocysteine (Hcy)-induced apoptosis in endothelial progenitor cells (EPCs). Methods Total mononuclear cells were isolated from mouse bone marrow by Ficoll density gradient centrifugation and were cultured in vitro for 7 d. Adherent cells were harvested and identified by fluorescence microscopy. EPCs were cultured with Hcy (0, 50, 100 and 500 μmol/L) for 12, 24 and 48 h, or pretreated with NAC(1 mmol/L), DPI(10 μmol/L) or SB203580 (10 μmol/L) for 30 min, then cultured with 500 μmol/L Hcy for 24 h. Apoptosis was detected by Annexin-V/PI flow cytometry, levels of reactive oxygen species (ROS) in cells were measured using H2DCF-DA as a fluorescence probe, NADPH oxidases were evaluated with lucigenin-enhanced chemiluminescence, and NO in the supernatant was determined by nitrate reductase assay. Results Hcy induced EPCs apoptosis, ROS accumulation, NADPH oxidase activation and decrease of NO in a time-dose dependent manner(P<0.05 or P<0.01). Pretreatment with NAC, DPI and SB203580 could inhibit these effects (P<0.05 or P<0.01). Conclusion Hcy could activate NADPH oxidase, induce ROS increase and NO decrease, and activate p38MAPK to enhance EPCs apoptosis.

Key words: homocysteine, endothelial progenitor cells, apoptosis, reactive oxygen species, NADPH oxidase, p38MAPK

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