Kitasato Institute for Life Sciences, Kitasato University, Minato-ku, Tokyo, Japan.
J Neuropathol Exp Neurol. 2010 Mar;69(3):224-33. doi: 10.1097/NEN.0b013e3181cfc46f.
The 50-kDa secreted glycoprotein pigment epithelium-derived factor (PEDF) is neuroprotective for various types of cultured neurons, but whether it is neuroprotective for neurons in vivo is not known. We examined the effects of adenovirus-mediated gene transfer of PEDF on quinolinic acid (QA)-induced neurotoxicity in rats. Adenoviral vector containing the human PEDF gene (Ad.PEDF) or Escherichia coli beta-galactosidase gene (Ad.LacZ) was directly injected into the right striatum 7 days before the injection of QA. Immunohistochemical analysis using antibodies specific for the neuronal markers dopamine and cyclic adenosine monophosphate-regulated phosphoprotein of 32 kDa, neuronal nuclei, and choline acetyltransferase revealed that the QA-induced striatal damage was significantly reduced in Ad.PEDF-treated rats. Overexpression of PEDF also reduced the expression of the inflammation-related genes for interleukin 1beta, tumor necrosis factor alpha, and macrophage inflammatory protein 1alpha 1 day after QA injection. Deletion analysis of human PEDF protein demonstrated that overexpression of PEDFDelta44-121 failed to protect neurons against QA-induced excitotoxicity, whereas PEDFDelta78-121 retained the neuroprotective activity, suggesting that amino acid residues 44-77 of the PEDF sequence are essential for PEDF-mediated neuroprotection in vivo. These results provide the first evidence that PEDF and its deletion mutant PEDFDelta78-121 are effective in protecting CNS neurons against excitotoxicity in vivo.
50kDa 分泌糖蛋白色素上皮衍生因子(PEDF)对各种培养神经元具有神经保护作用,但它是否对体内神经元具有神经保护作用尚不清楚。我们研究了腺病毒介导的 PEDF 基因转移对喹啉酸(QA)诱导的大鼠神经元毒性的影响。PEDF 基因(Ad.PEDF)或大肠杆菌β-半乳糖苷酶基因(Ad.LacZ)的腺病毒载体在 QA 注射前 7 天直接注射到右侧纹状体。使用针对神经元标志物多巴胺、环磷酸腺苷调节磷酸蛋白 32kDa、神经元核和胆碱乙酰转移酶的特异性抗体进行免疫组织化学分析表明,Ad.PEDF 处理大鼠的 QA 诱导的纹状体损伤明显减轻。PEDF 的过表达也降低了 QA 注射后 1 天炎症相关基因白细胞介素 1β、肿瘤坏死因子α和巨噬细胞炎症蛋白 1α的表达。人 PEDF 蛋白的缺失分析表明,PEDFDelta44-121 的过表达未能保护神经元免受 QA 诱导的兴奋性毒性,而 PEDFDelta78-121 保留了神经保护活性,表明 PEDF 序列的氨基酸残基 44-77 对于 PEDF 介导的体内神经保护是必需的。这些结果首次提供了证据,表明 PEDF 及其缺失突变体 PEDFDelta78-121 可有效保护中枢神经系统神经元免受体内兴奋性毒性。