title: Fentanyl activates hypoxia-inducible factor 1 in neuronal SH-SY5Y cells and mice under non-hypoxic conditions in a μ-opioid receptor-dependent manner.
abstract: Hypoxia-inducible factor 1 (HIF-1) is the main transcription factor responsible for hypoxia-induced gene expression. Perioperative drugs including anesthetics have been reported to affect HIF-1 activity. However, the effect of fentanyl on HIF-1 activity is not well documented. In this study, we investigated the effect of fentanyl and other opioids on HIF-1 activity in human SH-SY5Y neuroblastoma cells, hepatoma Hep3B cells, lung adenocarcinoma A549 cells and mice. Cells were exposed to fentanyl, and HIF-1 protein expression was examined by Western blot analysis using anti-HIF-1α and β antibodies. HIF-1-dependent gene expression was investigated by semi-quantitative real-time reverse transcriptase (RT)-PCR (qRT-PCR) and luciferase assay. Furthermore, fentanyl was administered intraperitoneally and HIF-1-dependent gene expression was investigated by qRT-PCR in the brains and kidneys of mice. A 10-μM concentration of fentanyl and other opioids, including 1 μM morphine and 4 μM remifentanil, induced HIF-1α protein expression and HIF-1 target gene expression in an opioid receptor-dependent manner in SH-SY5Y cells with activity peaking at 24h. Fentanyl did not augment HIF-1α expression during hypoxia-induced induction. HIF-1α stabilization assays and experiments with cycloheximide revealed that fentanyl increased translation from HIF-1α mRNA but did not stabilize the HIF-1α protein. Furthermore, fentanyl induced HIF-1 target gene expression in the brains of mice but not in their kidneys in a naloxone-sensitive manner. In this report, we describe for the first time that fentanyl, both in vitro and in vivo, induces HIF-1 activation under non-hypoxic conditions, leading to increases in expression of genes associated with adaptation to hypoxia.
authors.affiliation: Department of Anesthesia, Kyoto University Hospital, Kyoto, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Daijo
authors.forename: Shinichi
authors.initials: S
authors.lastname: Kai
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Tanaka
authors.forename: Takuhiko
authors.initials: T
authors.lastname: Wakamatsu
authors.forename: Shun
authors.initials: S
authors.lastname: Kishimoto
authors.forename: Kengo
authors.initials: K
authors.lastname: Suzuki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Harada
authors.forename: Satoshi
authors.initials: S
authors.lastname: Takabuchi
authors.forename: Takehiko
authors.initials: T
authors.lastname: Adachi
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Fukuda
authors.forename: Kiichi
authors.initials: K
authors.lastname: Hirota
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005283:Fentanyl / Q000008:administration & dosage / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051793:Hypoxia-Inducible Factor 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D009020:Morphine / Q000494:pharmacology; D009474:Neurons; D010880:Piperidines / Q000494:pharmacology; D055550:Protein Stability / Q000187:drug effects; D017450:Receptors, Opioid, mu / Q000378:metabolism*; D000077208:Remifentanil; D013995:Time
chemical_list: C497442:HIF1A protein, human; D051793:Hypoxia-Inducible Factor 1; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010880:Piperidines; D017450:Receptors, Opioid, mu; D009020:Morphine; D000077208:Remifentanil; D005283:Fentanyl