title: Differential in vitro actions of nitric oxide on human endometrial cell survival.
abstract: OBJECTIVE
  To investigate the presence of caspase-3 and Bcl-2 concentration in human endometrial tissue throughout the menstrual cycle, and study the effect of nitric oxide (NO) on cell proliferation and apoptosis during culture.
  
  
  METHODS
  Expression of caspase-3 and Bcl-2 concentration in endometrial explants, and examination of L-arginine (L-Arg) effect on epithelial and stromal cell proliferation and apoptosis in vitro.
  
  
  METHODS
  Prospective study.Twenty-seven eumenorrheic women (37 +/- 1.2 years).
  
  
  METHODS
  Endometrial samples were obtained with Pipelle suction curette from the corpus of the uterus.
  
  
  METHODS
  Apoptosis (annexin V-FITC binding), Bcl-2 concentration (ELISA), caspase-3 (immunohistochemistry), cell proliferation (spectrophotometric assay), and gene expression (RT-PCR).
  
  
  RESULTS
  Caspase-3 was detected by immunoassay in epithelial tissue throughout the menstrual cycle and in stroma during secretory phase. The Bcl-2 concentration was similar in endometrial homogenates obtained throughout the menstrual cycle, but L-Arg decreased Bcl-2 only in endometrium from the proliferative phase. In epithelial cells, NO increased apoptosis by 2.1 +/- 0.2-fold, augmented mRNA expression of Bax, and reduced expression of Bcl-2 compared with basal cultures. In stromal cells, NO increased cell proliferation in a dose-dependent manner, an effect that was blocked by a NO synthase inhibitor.
  
  
  CONCLUSIONS
  These data indicate that NO has a differential regulatory function on endometrial cell survival, as indicated by the results on stromal cell proliferation and epithelial cell apoptosis during culture, which suggests paracrine interactions between both cell types.
authors.affiliation: Institute of Maternal and Child Research, School of Medicine, University of Chile, Santiago, Chile. cjohnson@machi.med.uchile.cl
authors.forename: M Cecilia
authors.initials: MC
authors.lastname: Johnson
authors.forename: Manuel
authors.initials: M
authors.lastname: Maliqueo
authors.forename: M Angélica
authors.initials: MA
authors.lastname: Boric
authors.forename: Alejandra
authors.initials: A
authors.lastname: Villavicencio
authors.forename: David
authors.initials: D
authors.lastname: Vantman
authors.forename: Margarita
authors.initials: M
authors.lastname: Vega
mesh_terms: D000328:Adult; D017304:Annexin A5 / Q000032:analysis / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004717:Endometrium / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008597:Menstrual Cycle / Q000502:physiology*; D009569:Nitric Oxide / Q000494:pharmacology / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D017304:Annexin A5; C494826:BAX protein, human; D004791:Enzyme Inhibitors; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases