Tumor necrosis factor alpha as an endogenous stimulator for circulating coupling factor 6.

Abstract:

OBJECTIVE:We recently showed that mitochondrial coupling factor 6 (CF6) is present as a pressor substance and a prostacyclin inhibitor in systemic circulation. However, the regulation mechanism for circulating CF6 is unknown. We investigated the role of tumor necrosis factor-alpha (TNF-alpha) in the generation and release of CF6. METHODS AND RESULTS:We used two kinds of cells, human umbilical vein endothelial cells (HUVEC) and ECV-304. The concentration of CF6 in the medium increased with time in both ECV-304 and HUVEC. Treatment of ECV-304 and HUVEC with TNF-alpha enhanced the release of CF6 in a dose-dependent manner concomitantly with the decrease in CF6 content in the mitochondria at 24 h. The released CF6 was characterized to be an active full-length peptide by Western blot. The ratio of CF6 to GAPDH mRNA, measured by real-time polymerase chain reaction, was 1.7 fold increased at 1 h after exposure to TNF-alpha in ECV-304 and HUVEC. This enhanced gene expression and release was blocked or suppressed by 70% by stable transfection of dominant negative mutant I kappa B kinase alpha whose efficacy was confirmed by blockade of translocation of NF-kappa B p65 protein and of degradation of I kappa B alpha protein. Flow cytometry analysis revealed that the cell surface-associated CF6 was significantly increased at 24 h after TNF-alpha in a dose-dependent manner. CONCLUSIONS:TNF-alpha stimulates the gene expression of CF6 via activation of NF-kappa B signaling pathway, and promotes the release of CF6 from ECV-304 and HUVEC.

journal_name

Cardiovasc Res

journal_title

Cardiovascular research

authors

Sasaki S,Osanai T,Tomita H,Matsunaga T,Magota K,Okumura K

doi

10.1016/j.cardiores.2004.01.031

subject

Has Abstract

pub_date

2004-06-01 00:00:00

pages

578-86

issue

3

eissn

0008-6363

issn

1755-3245

pii

S0008636304000690

journal_volume

62

pub_type

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