Protection against ischemic brain injury by protein therapeutics
Protection against ischemic brain injury by protein therapeutics
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DOI：
10.1073/pnas.262460299
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发表时间：
2002-12-24
影响因子：
11.1
通讯作者：
Ohta, S
中科院分区：
文献类型：
--
作者：
Asoh, S;Ohsawa, I;Ohta, S
预防大量细胞死亡是各种损伤和疾病的重要治疗策略。蛋白质治疗剂具有在短时间内递送蛋白质的优点。我们已经设计了抗凋亡的bcl-x基因，以产生超级抗凋亡因子FNK，具有更强大的细胞保护活性。在这项研究中，我们融合了HIV/达特蛋白的蛋白转导结构域（PTD）FNK，并在缺血性脑损伤的动物模型中使用该构建体。将PTD-FNK加入人神经母细胞瘤细胞和大鼠新皮层神经元的培养液中，PTD-FNK迅速转导到细胞中，并在1 In内定位于线粒体。它分别保护神经母细胞瘤和神经元免于星形孢菌素诱导的细胞凋亡和谷氨酸诱导的兴奋性毒性。在低至0.3 pM的浓度下发现PTD-FNK的细胞保护活性。此外，PTD-FNK影响胞浆钙离子的运动，这可能与其神经保护作用有关。免疫组织化学分析显示，将myc标记的PTD-FNK（PTD-myc-FNK）腹腔注射到小鼠体内可以进入脑神经元。当腹腔注射到沙鼠，PTD-FNK防止在短暂的全脑缺血引起的海马迟发性神经元死亡。这些结果表明，PTD-FNK具有作为蛋白质治疗策略以防止脑中细胞死亡的临床效用的潜力。
Preventing massive cell death is an important therapeutic strategy for various injuries and disorders. Protein therapeutics have the advantage of delivering proteins in a short period. We have engineered the antiapoptotic bcl-x gene to generate the super antiapoptotic factor, FNK, with a more powerful cytoprotective activity. In this study, we fused the protein transduction domain (PTD) of the HIV/Tat protein to FNK and used the construct in an animal model of ischemic brain injury. When added into culture media of human neuroblastoma cells and rat neocortical neurons, PTD-FNK rapidly transduced into cells and localized to mitochondria within 1 In. it protected the neuroblastomas and neurons against staurosporine-induced apoptosis and glutamate-induced excitotoxicity, respectively. The cytoprotective activity of PTD-FNK was found at concentrations as low as 0.3 pM. Additionally, PTD-FNK affected the cytosolic movement of calcium ions, which may relate to its neuroprotective action. Immunohistochemical analysis revealed that myc-tagged PTD-FNK (PTD-myc-FNK) injected i.p. into mice can have access into brain neurons. When injected i.p. into gerbils, PTD-FNK prevented delayed neuronal death in the hippocampus caused by transient global ischemia. These results suggest that PTD-FNK has a potential for clinical utility as a protein therapeutic strategy to prevent cell death in the brain.