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Content archived on 2024-05-29

Regulation of apoptosis in fibroblast-like synoviocyte by TNF-alpha and signal transduction activation in the process

Objective

Hyperplasia of synovial membrane is a hallmark of Rheumatoid Arthritis (RA), and is associated with enhanced cytokine levels, which is also a character of RA. Synovial hyperplasia is mainly due to the fibroblast-like synoviocyte (FLS)'s over-proliferation and resistance to apoptosis. TNF-alpha is a primary cytokine in the development/progression of RA disease and is able to induce cell proliferation, activity and apoptosis. Clinicai experience has shown that anti-TNF-alpha therapy is effective without significant adverse effects. There has been some research in the field of TNF-alpha induced FLS proliferation, but few studies have been done about the regulation of apoptosis in FLS by TNF- alpha.

The aim of our research is to study regulation of apoptosis in FL S by TNF-alpha and the mechanism underlying this process. Apoptosis is an active action in cells determined by the signal transduction initiated by different signals such as cytokines. TNF-alpha is a cytokine broadly associated with both survival and death signal transduction pathways, and we hypothesize that TNF-alpha may play a dual role in FLS's apoptosis: on the one hand, it can protect FLS from apoptosis through activation of survival signal transduction such as PI3K-Akt signal pathway, and on the other hand, it also triggers the activation of some death signal transduction such as Fas signal pathway possibly leading to cells' apoptosis after the block of survival signal pathway.

TÚNEL (Terminal dUTP nick end labelling) assay and DNA laddering will be use d to identify apoptosis; cell viability will be determined by MTT assay; possible related signal factor expression will be measured by western-blot and RT-PCR on protein and mRNA level; electrophoretic mobility shift assay will be used to measure NF-kappa-B activity. Furthermore, using proteomic approach, we will explore factors involved in the regulation of TNF-alpha on apoptosis of FLS.

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Topic(s)

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Call for proposal

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FP6-2002-MOBILITY-7
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Funding Scheme

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IIF - Marie Curie actions-Incoming International Fellowships

Coordinator

AZIENDA OSPEDALIERA ARCISPEDALE SANTA MARIA NUOVA
EU contribution
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Total cost

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