title: Neuroprotective effect of TAT PTD-Ngb fusion protein on primary cortical neurons against hypoxia-induced apoptosis.
abstract: Hypoxic-ischemic injury increases neuroglobin (Ngb) expression in the brain. In our previous study, we have generated a transactivator-of-transcription protein-transduction domain-neuroglobin fusion protein (TAT PTD-Ngb) that successfully mediated exogenous Ngb expression in the primary neurons. In this study, we further investigated the role of TAT PTD-Ngb in protecting neurons against hypoxia-induced apoptosis and explored the possible mechanism. The primary cultured neurons were divided into four groups: (1) the normal group (no hypoxic injury); (2) the vehicle group (vehicle treatment and hypoxia injury); (3) the TAT PTD-Ngb group (TAT PTD-Ngb treatment and hypoxia injury); and (4) the Ngb group (Ngb treatment and hypoxia injury). Translocation of TAT PTD-Ngb into neurons was detected using fluorescent immunostaining against His-tag as early as 30 min after incubation. MTT assay showed that the TAT PTD-Ngb group had significantly increased cell viability compared to the vehicle or Ngb group after hypoxia. The result of transmission electron microscopy (TEM) also displayed rescued ultrastructure in TAT PTD-Ngb neurons compared to that of apoptotic neurons. In addition, TAT PTD-Ngb neurons showed significantly increased expression of anti-apoptotic Bcl-2 protein and decreased activities of caspase-3 and caspase-9 in response to hypoxia. These results suggest that TAT PTD-Ngb fusion protein protects primary cortical neurons against hypoxia-induced injury possibly through suppressing mitochondria apoptotic pathway.
authors: Department of Cadre Health Care, Qilu Hospital, Shandong University, Jinan, 250012, China.
authors: Guoyu
authors: G
authors: Zhou
authors: Peiyan
authors: P
authors: Shan
authors: Xueqiang
authors: X
authors: Hu
authors: Xueping
authors: X
authors: Zheng
authors: Shengnian
authors: S
authors: Zhou
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation / Q000187:drug effects; D015696:Gene Products, tat / Q000494:pharmacology*; D005914:Globins / Q000494:pharmacology*; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000494:pharmacology*; D000078787:Neuroglobin; D009474:Neurons / Q000187:drug effects* / Q000648:ultrastructure; D018696:Neuroprotective Agents / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D013997:Time Factors
chemical_list: D015696:Gene Products, tat; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D018696:Neuroprotective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D011993:Recombinant Fusion Proteins; D005914:Globins; D053148:Caspase 3; D053453:Caspase 9