title: Fluoxetine induces vascular endothelial growth factor/Netrin over-expression via the mediation of hypoxia-inducible factor 1-alpha in SH-SY5Y cells.
abstract: Fluoxetine has become one of the most promising drugs for improving clinical outcome in patients with cerebral infarction. Although the clinical efficacy of fluoxetine has been preliminarily demonstrated, its mechanism remains unclear. Hypoxia-inducible factor 1-alpha (HIF-1α) is upstream to Netrin and vascular endothelial growth factor (VEGF), and under hypoxia conditions it may induce expression of Netrin-1 and VEGF in vascular endothelial cells. We sought to explore whether it can regulate their expression in hypoxia and mediate the effect of fluoxetine in hypoxia. In this study, the effect of hypoxia on the expression of VEGF and Netrin was observed in vitro by real-time PCR and western blotting in SH-SY5Y cells; the binding sites of HIF-1α in VEGF and Netrin gene promoters were identified by luciferase reporter; the effect of fluoxetine on binding of HIF-1α with Netrin and VEGF promoters in hypoxia was observed by Chromatin Immunoprecipitation (ChIP) Assay. We prove that HIF-1α regulates transcription of both VEGF and Netrin, and that in hypoxia fluoxetine up-regulates VEGF and Netrin expression via mediation of HIF-1α that binds to hypoxia-response element sites of VEGF and Netrin promoters. Our study indicates that HIF-1α may play an important role in the treatment of cerebral infarction through mediating the recovery of neurological function induced by fluoxetine, which provides theoretical basis for the development of gene therapeutic drugs targeting HIF-1α. We show that hypoxia-inducible factor 1-alpha (HIF-1α) regulates transcription of both vascular endothelial growth factor (VEGF) and Netrin. Furthermore, we also show that in hypoxia fluoxetine up-regulates VEGF and Netrin expression via mediation of HIF-1α that binds to hypoxia-response element (HRE) sites of VEGF and Netrin promoters. Our study indicates that HIF-1α may play an important role in the treatment of cerebral infarction through mediating the recovery of neurological function induced by fluoxetine. These findings provide a theoretical basis for development of gene therapeutic drugs targeting HIF-1α.
authors.affiliation: Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Jiayi
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Mingrui
authors.initials: M
authors.lastname: Song
authors.affiliation: Department of Radiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Jinglong
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Neurology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Qiaoshu
authors.initials: Q
authors.lastname: Wang
keywords: fluoxetine; hypoxia-inducible factor 1-alpha; ischemic infarction; netrin; vascular endothelial growth factor