Mitochondrial Channel Opener Diazoxide Attenuates Hypoxia-Induced sFlt-1 Release in Human Choriocarcinoma Cells.
- Author:
Byeong Seop SHIN
1
;
Hwi Gon KIM
;
Ook Hwan CHOI
Author Information
- Publication Type:Original Article
- Keywords: Diazoxide; Heme oxygenase-1; Reactive oxygen speciese; Vascular endothelial growth factor receptor-1
- MeSH: Adenosine; Anoxia; Bilirubin; Blotting, Western; Choriocarcinoma*; Cobalt; Diazoxide*; Enzyme-Linked Immunosorbent Assay; Female; Heme Oxygenase-1; Humans; Polymerase Chain Reaction; Pregnancy; Reactive Oxygen Species; RNA, Messenger; RNA, Small Interfering; Tumor Necrosis Factor-alpha; Up-Regulation; Vascular Endothelial Growth Factor Receptor-1; Zinc
- From:Journal of Menopausal Medicine 2014;20(1):21-31
- CountryRepublic of Korea
- Language:English
- 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.
