title: EUK-207 protects human intestinal microvascular endothelial cells (HIMEC) against irradiation-induced apoptosis through the Bcl2 pathway.
abstract: OBJECTIVE
  To elucidate the signaling mechanisms involved in the protective effect of EUK-207 against irradiation-induced cellular damage and apoptosis in human intestinal microvasculature endothelial cells (HIMEC).
  
  
  METHODS
  HIMECs were irradiated and treated with EUK-207. Using hydroethidine and DCF-DA fluorescent probe the intracellular superoxide and reactive oxygen species (ROS) were determined. By real-time PCR and western blotting caspase-3, Bcl2 and Bax genes and proteins were analyzed. Proliferation was determined by [(3)H]-thymidine uptake. Immunofluorescence staining was used for translocation of p65 NFκB subunit.
  
  
  RESULTS
  Irradiation increased ROS production, apoptosis, Bax, Caspase3 and NFkB activity in HIMEC and inhibited cell survival/growth/proliferation. EUK-207 restored the endothelial functions, markedly inhibited the ROS, up-regulated the Bcl2 and down-regulated Bax and prevented NFκB caspase 3 activity in HIMEC.
  
  
  CONCLUSIONS
  HIMEC provide a novel model to define the effect of irradiation induced endothelial dysfunction. Our findings suggest that EUK-207 effectively inhibits the damaging effect of irradiation.
authors.affiliation: Department of Surgery, Medical College of Wisconsin, Milwaukee, WI 53226, United States.
authors.forename: Mary F
authors.initials: MF
authors.lastname: Otterson
authors.forename: Linghui
authors.initials: L
authors.lastname: Nie
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Schmidt
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Link
authors.forename: Nebojsa
authors.initials: N
authors.lastname: Jovanovic
authors.forename: Orestis
authors.initials: O
authors.lastname: Lyros
authors.forename: Parvaneh
authors.initials: P
authors.lastname: Rafiee
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000528:radiation effects; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D019610:Cytoprotection / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects*; D016166:Free Radical Scavengers / Q000494:pharmacology*; D006801:Humans; D007422:Intestines / Q000098:blood supply; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008315:Malondialdehyde / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009942:Organometallic Compounds / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000528:radiation effects; D013482:Superoxide Dismutase / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: C476761:EUK 207; D016166:Free Radical Scavengers; D016328:NF-kappa B; D009942:Organometallic Compounds; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D008315:Malondialdehyde; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase; D053148:Caspase 3