Tumor necrosis factor-alpha-induced nuclear factor-kappaB activation in human cardiomyocytes is mediated by NADPH oxidase
J. physiol. biochem
; 70(3): 769-779, sept. 2014.
Article
in En
| IBECS
| ID: ibc-127321
Responsible library:
ES1.1
Localization: BNCS
ABSTRACT
An elevated level of tumor necrosis factor (TNF)-α is implicated in several cardiovascular diseases including heart failure. Numerous reports have demonstrated that TNF- alfa activates nuclear factor (NF)-kappaB, resulting in the upregulation of several genes that regulate inflammation, proliferation, and apoptosis of cardiomyocytes. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, a major source of reactive oxygen species (ROS), is also activated by TNF- alfa and plays a crucial role in redox-sensitive signaling pathways. The present study investigated whether NADPH oxidase mediates TNF-alfa-induced NF-kappaB activation and NF-kappaB-mediated gene expression. Human cardiomyocytes were treated with recombinant TNF- alfa with or without pretreatment with diphenyleneiodonium (DPI) and apocynin, inhibitors of NADPH oxidase. TNF-α-induced ROS production was measured using 5-(and-6)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate assay. TNF- alfa-induced NF-kappaB activation was also examined using immunoblot; NF-kappaB binding to its binding motif was determined using a Cignal reporter luciferase assay and an electrophoretic mobility shift assay. TNF- alfa -induced upregulation of interleukin (IL)-1β and vascular cell adhesion molecule (VCAM)-1 was investigated using real-time PCR and immunoblot. TNF- alfa -induced ROS production in cardiomyocytes was mediated by NADPH oxidase. Phosphorylation of IKK- alfa /β and p65, degradation of IkappaBalfa, binding of NF-kappaB to its binding motif, and upregulation of IL-1β and VCAM-1 induced by TNF- alfa were significantly attenuated by treatment with DPI and apocynin. Collectively, these findings demonstrate that NADPH oxidase plays a role in regulation of TNF- alfa -induced NF-kappaB activation and upregulation of proinflammatory cytokines, IL-1β and VCAM-1, in human cardiomyocytes
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Collection:
06-national
/
ES
Database:
IBECS
Main subject:
Tumor Necrosis Factor-alpha
/
Myocytes, Cardiac
/
Receptor Activator of Nuclear Factor-kappa B
Limits:
Humans
Language:
En
Journal:
J. physiol. biochem
Year:
2014
Document type:
Article