title: Combining Normobaric Oxygen with Ethanol or Hypothermia Prevents Brain Damage from Thromboembolic Stroke via PKC-Akt-NOX Modulation.
abstract: In a thromboembolic stroke model after reperfusion by recombinant tissue plasminogen activator (rt-PA), we aimed to determine whether therapeutic hypothermia (TH) and ethanol (EtOH) in combination with low concentration (60 %) of normobaric oxygen (NBO) enhanced neuroprotection, as compared to using each of these agents alone. We further aimed to elucidate a potential role of the NADPH oxidase (NOX), phosphorylated protein kinase B (Akt), and protein kinase C-δ (PKC-δ) pathway in oxidative stress and neuroprotection. In Sprague-Dawley rats, a focal middle cerebral artery (MCA) occlusion was induced by an autologous embolus in the following experimental groups: rt-PA treatment alone, rt-PA + NBO treatment, rt-PA + TH at 33 °C, rt-PA + EtOH, rt-PA + NBO + EtOH, rt-PA + NBO + TH, rt-PA + NOX inhibitor, rt-PA + EtOH + NOX inhibitor, or rt-PA + EtOH + Akt inhibitor. Control groups included sham-operated without stroke or stroke without treatment. Infarct volume and neurological deficit were assessed at 24 h after rt-PA-induced reperfusion with or without treatments. ROS levels, NOX activity, and the protein expression of NOX subunits p22phox, p47phox, p67phox, gp91phox, as well as PKC-δ and phosphorylated Akt were measured at 3 and 24 h after rt-PA-induced reperfusion. Following rt-PA in thromboembolic stroke rats, NBO combined with TH or EtOH more effectively decreased infarct volume and neurological deficit, as well as reactive oxygen species (ROS) production than with any of the used monotherapies. NOX activity and subunit expressions were downregulated and temporally associated with reduced PKC-δ and increased p-Akt expression. The present study demonstrated that combining NBO with either TH or EtOH conferred similar neuroprotection via modulation of NOX activation. The results suggest a role of Akt in NOX activation and implicate an upstream PKC-δ pathway in the Akt regulation of NOX. It is possible to substitute EtOH for TH, thus circumventing the difficulties in clinical application of TH through the comparatively easier usage of EtOH as a potential stroke management.
authors.affiliation: China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing, 101149, China.|Department of Neurology, Luhe Hospital, Capital Medical University, Beijing, China.|Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: Lipeng
authors.initials: L
authors.lastname: Cai
authors.affiliation: Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: James
authors.initials: J
authors.lastname: Stevenson
authors.affiliation: Department of Neurology, Luhe Hospital, Capital Medical University, Beijing, China.|Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Geng
authors.affiliation: Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: Changya
authors.initials: C
authors.lastname: Peng
authors.affiliation: China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing, 101149, China. jixm@ccmu.edu.cn.|Department of Neurology, Luhe Hospital, Capital Medical University, Beijing, China. jixm@ccmu.edu.cn.
authors.forename: Xunming
authors.initials: X
authors.lastname: Ji
authors.affiliation: Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.|Department of Radiology, Luhe Hospital, Capital Medical University, Beijing, China.
authors.forename: Ruiqiang
authors.initials: R
authors.lastname: Xin
authors.affiliation: Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Rastogi
authors.affiliation: Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Sy
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Rafols
authors.affiliation: China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing, 101149, China. yding@med.wayne.edu.|Department of Neurology, Luhe Hospital, Capital Medical University, Beijing, China. yding@med.wayne.edu.|Department of Neurological Surgery, Wayne State University School of Medicine, 550 E Canfield, Detroit, MI, 48201, USA. yding@med.wayne.edu.
authors.forename: Yuchuan
authors.initials: Y
authors.lastname: Ding
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D001930:Brain Injuries / Q000378:metabolism / Q000517:prevention & control*; D003131:Combined Modality Therapy / Q000379:methods; D000431:Ethanol / Q000008:administration & dosage*; D007036:Hypothermia, Induced / Q000379:methods*; D019255:NADPH Oxidases / Q000502:physiology; D010100:Oxygen / Q000008:administration & dosage*; D051745:Protein Kinase C-delta / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D020521:Stroke / Q000378:metabolism / Q000628:therapy*; D013923:Thromboembolism / Q000378:metabolism / Q000628:therapy*
chemical_list: D000431:Ethanol; D019255:NADPH Oxidases; D051057:Proto-Oncogene Proteins c-akt; D051745:Protein Kinase C-delta; D010100:Oxygen
keywords: Akt; Autologous embolus; Ischemia/reperfusion; NOX; Neuroprotection; PKC-δ; ROS; Thrombolytic therapy
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