title: Hypoxia-inducible factor 1 contributes to N-acetylcysteine's protection in stroke.
abstract: Stroke is a leading cause of adult morbidity and mortality with very limited treatment options. Evidence from preclinical models of ischemic stroke has demonstrated that the antioxidant N-acetylcysteine (NAC) effectively protects the brain from ischemic injury. Here, we evaluated a new pathway through which NAC exerted its neuroprotection in a transient cerebral ischemia animal model. Our results demonstrated that pretreatment with NAC increased protein levels of hypoxia-inducible factor-1α (HIF-1α), the regulatable subunit of HIF-1, and its target proteins erythropoietin (EPO) and glucose transporter (GLUT)-3, in the ipsilateral hemispheres of rodents subjected to 90min middle cerebral artery occlusion (MCAO) and 24h reperfusion. Interestingly, after NAC pretreatment and stroke, the contralateral hemisphere also demonstrated increased levels of HIF-1α, EPO, and GLUT-3, but to a lesser extent. Suppressing HIF-1 activity with two widely used pharmacological inhibitors, YC-1 and 2ME2, and specific knockout of neuronal HIF-1α abolished NAC's neuroprotective effects. The results also showed that YC-1 and 2ME2 massively enlarged infarcts, indicating that their toxic effect was larger than just abolishing NAC's neuroprotective effects. Furthermore, we determined the mechanism of NAC-mediated HIF-1α induction. We observed that NAC pretreatment upregulated heat-shock protein 90 (Hsp90) expression and increased the interaction of Hsp90 with HIF-1α in ischemic brains. The enhanced association of Hsp90 with HIF-1α increased HIF-1α stability. Moreover, Hsp90 inhibition attenuated NAC-induced HIF-1α protein accumulation and diminished NAC-induced neuroprotection in the MCAO model. These results strongly indicate that HIF-1 plays an important role in NAC-mediated neuroprotection and provide a new molecular mechanism involved in the antioxidant's neuroprotection in ischemic stroke.
authors.affiliation: Department of Pharmacology & Toxicology, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: Ziyan
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology & Toxicology, University of Kansas, Lawrence, KS 66045, USA.
authors.forename: Jingqi
authors.initials: J
authors.lastname: Yan
authors.affiliation: Department of Radiology, Medical University of South Carolina, Charleston, SC 29401, USA.
authors.forename: Saeid
authors.initials: S
authors.lastname: Taheri
authors.affiliation: Department of Pharmaceutical Sciences, University of New Mexico, Albuquerque, NM 87131, USA.
authors.forename: Ke Jian
authors.initials: KJ
authors.lastname: Liu
authors.affiliation: Department of Pharmacology & Toxicology, University of Kansas, Lawrence, KS 66045, USA. Electronic address: hshi@ku.edu.
authors.forename: Honglian
authors.initials: H
authors.lastname: Shi
mesh_terms: D000111:Acetylcysteine / Q000008:administration & dosage* / Q000378:metabolism; D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D020244:Infarction, Middle Cerebral Artery / Q000378:metabolism / Q000473:pathology*; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000378:metabolism; D051381:Rats; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction; D020521:Stroke / Q000378:metabolism* / Q000473:pathology
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D018696:Neuroprotective Agents; D000111:Acetylcysteine
keywords: Free radicals; HIF-1; Hsp90; NAC; Neuroprotection; Stroke