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Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells

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% O(2)) or hypoxia (1% O(2)), and expression of sFlt-1 mRNA and protein release was determined by quanti...

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Autores principales: Shin, Byeong Seop, Kim, Hwi Gon, Choi, Ook Hwan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society of Menopause 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4217563/
https://www.ncbi.nlm.nih.gov/pubmed/25371888
http://dx.doi.org/10.6118/jmm.2014.20.1.21
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author Shin, Byeong Seop
Kim, Hwi Gon
Choi, Ook Hwan
author_facet Shin, Byeong Seop
Kim, Hwi Gon
Choi, Ook Hwan
author_sort Shin, Byeong Seop
collection PubMed
description 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% O(2)) or hypoxia (1% O(2)), 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-α) 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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spelling pubmed-42175632014-11-04 Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells Shin, Byeong Seop Kim, Hwi Gon Choi, Ook Hwan J Menopausal Med Original Article 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% O(2)) or hypoxia (1% O(2)), 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-α) 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. The Korean Society of Menopause 2014-04 2014-04-28 /pmc/articles/PMC4217563/ /pubmed/25371888 http://dx.doi.org/10.6118/jmm.2014.20.1.21 Text en Copyright © 2014 by The Korean Society of Menopause http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/).
spellingShingle Original Article
Shin, Byeong Seop
Kim, Hwi Gon
Choi, Ook Hwan
Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title_full Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title_fullStr Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title_full_unstemmed Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title_short Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells
title_sort mitochondrial channel opener diazoxide attenuates hypoxia-induced sflt-1 release in human choriocarcinoma cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4217563/
https://www.ncbi.nlm.nih.gov/pubmed/25371888
http://dx.doi.org/10.6118/jmm.2014.20.1.21
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