title: Evidence that peroxynitrite affects human osteoblast proliferation and differentiation.
abstract: Peroxynitrite (PN), a nitric oxide (NO*)-derived anion, has been associated with NO* damage in various cell types. We examined the effects of adding PN to cultured human osteoblast-like (hOB) cells obtained after hip arthroplasty. Exposure to PN (0.1-0.4 mM) decreased both hOB proliferation and differentiation, measured by [3H]thymidine uptake and alkaline phosphatase production, respectively. Incubation with 3-morpholinosydnonimine (SIN-1; 0.25-1 mM), an NO* and O2- donor that leads to PN release, also reduced both hOB proliferation and differentiation. Coincubation with both superoxide dismutase (SOD; 100 U/ml) and catalase (CAT; 50 U/ml), rendering SIN-1 a pure NO* donor, reversed its effects on hOB proliferation and differentiation. However, SIN-1-induced NO* production, measured by nitrite release to the hOB medium, was not altered by cotreatment with SOD and CAT. Expression of nitrotyrosine by hOB, a marker of PN action, was significantly increased after SIN-1 addition, as compared with untreated cells, as revealed by Western blot analysis. Interleukin-1alpha (IL-1alpha) and interferon gamma (IFN-gamma) but not tumor necrosis factor alpha (TNF-alpha) also significantly increased nitrotyrosine expression in these cells. These data show that PN is at least partially responsible for osteoblast derangement by NO* and that cytokines released during inflammatory arthropathies can induce PN production in hOB cells.
authors.affiliation: Department of Medicine, Faculty of Medicine, Université de Sherbrooke, Québec, Canada.
authors.forename: Francisco Airton Castro
authors.initials: FA
authors.lastname: da Rocha
authors.forename: Artur José
authors.initials: AJ
authors.lastname: de Brum-Fernandes
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis; D002374:Catalase / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009573:Nitrites / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins; D013482:Superoxide Dismutase / Q000494:pharmacology; D013936:Thymidine / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism
chemical_list: D007375:Interleukin-1; D020030:Nitric Oxide Donors; D009573:Nitrites; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D030421:Peroxynitrous Acid; D014443:Tyrosine; C002385:linsidomine; D007371:Interferon-gamma; D008981:Molsidomine; D002374:Catalase; D013482:Superoxide Dismutase; D000469:Alkaline Phosphatase; D013936:Thymidine