title: TAT-mediated delivery of neuroglobin protects against focal cerebral ischemia in mice.
abstract: Neuroglobin (Ngb), a newly identified globin in vertebrate brain, has been suggested to be able to protect against brain hypoxic-ischemic injury. However, owing to its large size, the impermeability of the blood-brain barrier (BBB) to Ngb limits its application in brain injury. Recently, the 11-amino-acid human immunodeficiency virus trans-activator of transcription (TAT) protein transduction domain was shown to successfully deliver macromolecules into the brain. This study explored whether the TAT-Ngb fusion protein can cross the BBB and protect the brain from cerebral ischemia. The TAT-Ngb fusion protein generated from Escherichia coli BL21 (DE3) was efficiently delivered into mice brain tissues by intravenous injection as demonstrated by immunohistochemistry and Western blotting. Two groups of mice were treated with filamentous middle cerebral artery occlusion (MCAO) for 30min or 2h followed by reperfusion. Each group was then divided into sub-groups and was injected intravenously with TAT-Ngb, Ngb, or saline respectively before MCAO or immediately after reperfusion. Compared with the Ngb- and saline-treated group, the group with TAT-Ngb treated 2h before MCAO showed significantly less brain infarct volume and had better neurologic outcomes (p<0.05). Furthermore, a TAT-Ngb injection following a 30-min MCAO treatment significantly increased neuronal survival in the striatum (p<0.05). Our results demonstrated that the exogenous Ngb fusion protein containing the TAT protein transduction domain could be efficiently transduced into neurons in the mouse and protect the brain from mild or moderate ischemic injury.
authors.affiliation: Department of Neurology and Institute of Neurology, First Affiliated Hospital, Center of Neuroscience, Fujian Medical University, 20 Chazhong Road, Fuzhou 350005, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Cai
authors.forename: Yi
authors.initials: Y
authors.lastname: Lin
authors.forename: Xie-Hua
authors.initials: XH
authors.lastname: Xue
authors.forename: Ling
authors.initials: L
authors.lastname: Fang
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Zhi-Ying
authors.initials: ZY
authors.lastname: Wu
mesh_terms: D000818:Animals; D001784:Blood Gas Analysis / Q000379:methods; D001794:Blood Pressure / Q000502:physiology; D020520:Brain Infarction / Q000209:etiology / Q000517:prevention & control; D004195:Disease Models, Animal; D005822:Genetic Vectors / Q000502:physiology; D005914:Globins / Q000008:administration & dosage*; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods; D020244:Infarction, Middle Cerebral Artery / Q000150:complications / Q000517:prevention & control*; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000008:administration & dosage*; D009422:Nervous System Diseases / Q000209:etiology / Q000517:prevention & control; D000078787:Neuroglobin; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics; D012039:Regional Blood Flow / Q000502:physiology; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*
chemical_list: D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D005914:Globins; D010751:Phosphopyruvate Hydratase