Tumor necrosis factor-α protects synovial cells from nitric oxide induced apoptosis through phosphoinositide 3-kinase Akt signal transduction

Qingquan Chen, Bruno Casali, Laura Pattacini, Luigi Boiardi, Carlo Salvarani

Research output: Contribution to journalArticle

Abstract

Objective. To investigate the anti-apoptotic role of tumor necrosis factor-α (TNF-α) 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-α treatment prevented FLS apoptosis induced by nitric oxide (NO) donor sodium nitroprusside dihydrate (SNP), achieving 70% protection. At 1-10 ng·ml -1 concentrations, TNF-α 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-κB inhibitor pyrrolidine-dithiocarbamate. Conclusion. TNF-α has an anti-apoptotic effect in human FLS. Activation of Akt and BAD may have an important role in this process.

Original languageEnglish
Pages (from-to)1061-1068
Number of pages8
JournalJournal of Rheumatology
Volume33
Issue number6
Publication statusPublished - Jun 2006

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1-Phosphatidylinositol 4-Kinase
Signal Transduction
Nitric Oxide
Tumor Necrosis Factor-alpha
Fibroblasts
Apoptosis
Eagles
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Nitric Oxide Donors
In Situ Nick-End Labeling
Nitroprusside
Rheumatoid Arthritis
Cell Survival
Western Blotting
Phosphorylation
Staining and Labeling
Synoviocytes
Therapeutics

Keywords

  • Apoptosis
  • Nitric oxide
  • Signal transduction
  • Synovial cells
  • Tumor necrosis factor-α

ASJC Scopus subject areas

  • Rheumatology
  • Immunology

Cite this

Tumor necrosis factor-α protects synovial cells from nitric oxide induced apoptosis through phosphoinositide 3-kinase Akt signal transduction. / Chen, Qingquan; Casali, Bruno; Pattacini, Laura; Boiardi, Luigi; Salvarani, Carlo.

In: Journal of Rheumatology, Vol. 33, No. 6, 06.2006, p. 1061-1068.

Research output: Contribution to journalArticle

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AU - Chen, Qingquan

AU - Casali, Bruno

AU - Pattacini, Laura

AU - Boiardi, Luigi

AU - Salvarani, Carlo

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N2 - Objective. To investigate the anti-apoptotic role of tumor necrosis factor-α (TNF-α) 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-α treatment prevented FLS apoptosis induced by nitric oxide (NO) donor sodium nitroprusside dihydrate (SNP), achieving 70% protection. At 1-10 ng·ml -1 concentrations, TNF-α 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-κB inhibitor pyrrolidine-dithiocarbamate. Conclusion. TNF-α has an anti-apoptotic effect in human FLS. Activation of Akt and BAD may have an important role in this process.

AB - Objective. To investigate the anti-apoptotic role of tumor necrosis factor-α (TNF-α) 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-α treatment prevented FLS apoptosis induced by nitric oxide (NO) donor sodium nitroprusside dihydrate (SNP), achieving 70% protection. At 1-10 ng·ml -1 concentrations, TNF-α 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-κB inhibitor pyrrolidine-dithiocarbamate. Conclusion. TNF-α has an anti-apoptotic effect in human FLS. Activation of Akt and BAD may have an important role in this process.

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