title: Tumor necrosis factor-alpha protects synovial cells from nitric oxide induced apoptosis through phosphoinositide 3-kinase Akt signal transduction.
abstract: OBJECTIVE
  To investigate the anti-apoptotic role of tumor necrosis factor-a (TNF-a) and its signaling pathways in cultured human fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis.
  
  
  METHODS
  FLS were cultured in Dulbecco's modified Eagle's medium. Apoptotic cells were identified by TUNEL assay and Hoechst staining. Cell viability was determined by the MTT method. Expression of phospho-Akt and phospho-BAD was measured by Western blotting.
  
  
  RESULTS
  A 24-h TNF-a treatment prevented FLS apoptosis induced by nitric oxide (NO) donor sodium nitroprusside dihydrate (SNP), achieving 70% protection. At 1-10 ng x ml-1 concentrations, TNF-a induced phosphorylation of Akt and BAD in a time and concentration-dependent manner. This effect was blocked by treatment with both LY294002 and nuclear factor-kB inhibitor pyrrolidine-dithiocarbamate.
  
  
  CONCLUSIONS
  TNF-a has an anti-apoptotic effect in human FLS. Activation of Akt and BAD may have an important role in this process.
authors.affiliation: Laboratorio di Biologia Molecolare and Unità di Reumatologia, Arcispedale S. Maria Nuova, Reggio Emilia, Italy.
authors.forename: Qingquan
authors.initials: Q
authors.lastname: Chen
authors.forename: Bruno
authors.initials: B
authors.lastname: Casali
authors.forename: Laura
authors.initials: L
authors.lastname: Pattacini
authors.forename: Luigi
authors.initials: L
authors.lastname: Boiardi
authors.forename: Carlo
authors.initials: C
authors.lastname: Salvarani
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004336:Drug Antagonism; D004338:Drug Combinations; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009569:Nitric Oxide / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D011759:Pyrrolidines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013583:Synovial Membrane / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013859:Thiocarbamates / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D051025:bcl-Associated Death Protein / Q000378:metabolism
chemical_list: D002867:Chromones; D004338:Drug Combinations; D009025:Morpholines; D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; D051025:bcl-Associated Death Protein; D009599:Nitroprusside; C020972:pyrrolidine dithiocarbamic acid; D009569:Nitric Oxide; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D019869:Phosphatidylinositol 3-Kinases; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt