Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
Journal of Menopausal Medicine
; : 21-31, 2014.
Article
em En
| WPRIM
| ID: wpr-228702
Biblioteca responsável:
WPRO
ABSTRACT
OBJECTIVES: To examine the effect of diazoxide on hypoxia-induced soluble fms-like tyrosin kinase-1 (sFlt-1) release in JEG-3 choriocarcinoma cells. METHODS: Cells were cultured under normoxia (20% O2) or hypoxia (1% O2), and expression of sFlt-1 mRNA and protein release was determined by quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) assays and enzyme-linked immunosorbent assay (ELISA). RESULTS: Tumor necrosis factor-alpha (TNF-alpha) as well as hypoxia stimulated sFlt-1 release and diazoxide inhibited both of them. The selective inhibitor of mitochondrial adenosine triphosphat (ATP)-sensitive K+ channel opener (K(ATP)) 5-hydroxydecanoate (5-HD) completely reversed the diazoxide-induced inhibition of hypoxia-stimulated sFlt-1 release. qRT-PCR and Western blot analyses showed that diazoxide up-regulated the heme oxygenase-1 (HO-1) expression. In addition, the HO-1 inducer cobalt protoporphyrin (CoPP) and the metabolic product of HO-1 bilirubin mimicked diazoxide to inhibit sFlt-1 release and reactive oxygen species (ROS) production under hypoxia, whereas the HO-1 inhibitor zinc protoporphyrin IX (ZnPP IX) antagonized the effect of diazoxide. In cells transfected with the HO-1 siRNA, diazoxide did not exert any effect on sFlt-1 release and ROS production under hypoxia. CONCLUSION: These results, taken together, strongly suggest that up-regulation of the HO-1 expression is the crucial mechanism responsible for the diazoxide-induced inhibition of the sFlt-1 release and ROS production under hypoxia.
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Base de dados:
WPRIM
Assunto principal:
Zinco
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Bilirrubina
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RNA Mensageiro
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Ensaio de Imunoadsorção Enzimática
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Coriocarcinoma
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Adenosina
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Regulação para Cima
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Western Blotting
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Reação em Cadeia da Polimerase
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Fator de Necrose Tumoral alfa
Limite:
Female
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Humans
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Pregnancy
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article