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J Rheumatol. 2006 Jun;33(6):1061-8.

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

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1
Laboratorio di Biologia Molecolare and Unità di Reumatologia, Arcispedale S. Maria Nuova, Reggio Emilia, Italy.

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.

CONCLUSION:

TNF-a has an anti-apoptotic effect in human FLS. Activation of Akt and BAD may have an important role in this process.

PMID:
16755653
[Indexed for MEDLINE]
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