The intravenous anesthetics barbiturates inhibit hypoxia-inducible factor 1 activation.
Hypoxia-inducible factor 1 (HIF-1) is a master transcription factor of hypoxia-induced gene expression. Anesthetics and perioperative drugs have been reported to affect HIF-1 activity. However, the effect of barbiturates on HIF-1 activity has not been reported. In this study, we investigated the effect of thiopental and thiamylal on HIF-1 activity using the neuronal SH-SY5Y cells, the non-neuronal HEK293 cells, and the macrophage-differentiated THP-1 cells. Cells were exposed to 20% or 1% O(2) conditions with or without thiopental or thiamylal treatment. The cell lysate were subjected to Western blot analysis using anti-HIF-1alpha and -HIF-1beta antibodies. HIF-1-dependent gene expression was investigated by semi-quantitative real-time RT-PCR and luciferase assay. Hydroxylation of HIF-1alpha protein was evaluated by in vitro pulldown assay using recombinant protein. Both thiopental and thiamylal reversibly suppressed hypoxia-induced HIF-1 activation in the neuronal and the non-neuronal cells in a dose-dependent manner. Moreover, the barbiturates inhibited lipopolysaccharide-induced HIF-1alpha expression in THP-1 cells. The HIF-1-downstream gene expression was also inhibited by the barbiturates. HIFalpha-hydroxylases activity and HIF-1alpha stability were not affected but the HIF-1alpha protein neosynthesis was inhibited by the barbiturates. Our experimental results indicate that barbiturates inhibit induced HIF-1 activation and downstream genes expression.
Wakamatsu
Takuhiko
T
Department of Anesthesia, Kyoto University Hospital, Kyoto University, Kyoto, Japan.
Tanaka
Tomoharu
T
Oda
Seiko
S
Nishi
Kenichiro
K
Harada
Hiroshi
H
Daijo
Hiroki
H
Takabuchi
Satoshi
S
Kai
Shinichi
S
Fukuda
Kazuhiko
K
Hirota
Kiichi
K
D000771:Anesthesia, Intravenous; D000777:Anesthetics / Q000008:administration & dosage / Q000494:pharmacology*; D000818:Animals; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D001463:Barbiturates / Q000008:administration & dosage / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D006801:Humans; D051793:Hypoxia-Inducible Factor 1 / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D015533:Transcriptional Activation / Q000187:drug effects*
C497396:ARNT protein, human; D000777:Anesthetics; D001463:Barbiturates; C497442:HIF1A protein, human; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D010100:Oxygen; C032232:barbituric acid