title: Atorvastatin attenuates homocysteine-induced apoptosis in human umbilical vein endothelial cells via inhibiting NADPH oxidase-related oxidative stress-triggered p38MAPK signaling.
abstract: OBJECTIVE
  To examine the effect of atorvastatin on homocysteine (Hcy)-induced reactive oxygen species (ROS) production and apoptosis in human umbilical vein endothelial cells (HUVECs).
  
  
  METHODS
  HUVECs were cultured with Hcy (0.1-5 mmol/L) in the presence or absence of atorvastatin (1-100 micromol//L) or various stress signaling inhibitors, including the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium (DPI, 10 micromol/L), the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580 (10 micromol/L) and antioxidants N-acetyl cysteine (NAC, 1 mmol/L). Cell apoptosis was evaluated by Annexin V/propidium iodide staining and flow cytometry. ROS were detected by 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFH-DA). NADPH oxidases were evaluated with lucigenin-enhanced chemiluminescence. Hcy-induced expression of p38MAPK protein was measured by Western blotting analysis.
  
  
  RESULTS
  Atorvastatin inhibited endothelial cell apoptosis induced by 1 mmol/L Hcy in a dose-dependent manner and the maximal inhibitory effect was reached at 100 micromol/L. Atorvastatin (10 micromol/L) significantly suppressed Hcy (1 mmol/L for 30 min) induced ROS accumulation (3.17+/-0.33 vs 4.34+/-0.31, P<0.05). Atorvastatin (10 micromol/L) also antagonized Hcy (1 mmol/L for 30 min) induced activation of NADPH oxidase (2.57+/-0.49 vs 3.33+/-0.6, P<0.05). Furthermore, atorvastatin inhibited Hcy-induced phosphorylation of p38 MAPK (1.7+/-0.1 vs 2.22+/-0.25, P<0.05), similar effects occurred with DPI, NAC and SB203580.
  
  
  CONCLUSIONS
  Atorvastatin may inhibit Hcy-induced ROS accumulation and endothelium cell apoptosis through an NADPH oxidase and/or p38MAPK-dependent mechanisms, all of which may contribute to atorvastatin-induced beneficial effect on endothelial function.
authors.affiliation: Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Xiao-mei
authors.initials: XM
authors.lastname: Bao
authors.forename: Chun-fang
authors.initials: CF
authors.lastname: Wu
authors.forename: Guo-ping
authors.initials: GP
authors.lastname: Lu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D000069059:Atorvastatin; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006538:Heptanoic Acids / Q000494:pharmacology*; D006710:Homocysteine / Q000494:pharmacology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism; D009861:Onium Compounds / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D011725:Pyridines / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000032:analysis / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D006538:Heptanoic Acids; D007093:Imidazoles; D009861:Onium Compounds; D011725:Pyridines; D011758:Pyrroles; D017382:Reactive Oxygen Species; D006710:Homocysteine; C007517:diphenyleneiodonium; D000069059:Atorvastatin; D019255:NADPH Oxidases; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580