An adenovirus vector for neurotrophin transfection induces compressed spinal cord of adults rat

用于神经营养素转染的腺病毒载体诱导成年大鼠脊髓受压

• 批准号: 12470305
• 负责人: BABA Hisatoshi
• 金额: $ 7.23万
• 依托单位: Fukui Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470305/
• 关键词: adenovirus vector; neurotrophin; compressed spinal cord; adult rat; 遺伝子治療; ウイルスベクター

We investigated neurotrotective effect of glial cell lined-derived neurotrophic factor (GDNF) encoded by an adenovirus vector (AxCAhGDNF) on the injured spinal cord lesions. The rat thoracic spinal cord was experimentally injured by extradural static weight compression. After the direct injection of the adenovirus vector into thoracic spinal cord, histological and immunohistochemical analysis and immunoblot analysis were examined. One week after the injury with AxCAlacZ, the animals showed expression of β-galactosidase activity in injured and injected lesions and the expression was visualized until 4-6 weeks after injection. Animals injured and injected with AxCAhGDNF showed intense immunolabeling for GDNF in lesioned spinal cord motoneurons, reactive astrocyte and oligodendrocyte, and expression of virus induced human GDNF mRNA transcripts in the spinal cord tissue. Nissl-stained cell counts revealed that the injection with AxCAhGDNF significantly prevented the loss of lesioned spinal cord motoneurons 2-4 weeks after injury. These results indicate that the adenovirus-mediated gene transfer of GDNF may prevent the degeneration of motoneurons in adult humans with spinal cord injury.

目的：研究腺病毒载体介导的胶质细胞源性神经营养因子（glialcelllined-derivedneurotrophicfactor，GDNF）对脊髓损伤的保护作用。用静态加压法造成大鼠胸段脊髓损伤。将腺病毒载体直接注入胸段脊髓后，进行组织学、免疫组化和免疫印迹分析。在用AxCAlacZ损伤后一周，动物在损伤的和注射的病变中显示β-半乳糖苷酶活性的表达，并且该表达直到注射后4-6周才显现。动物损伤和AxCAhGDNF注射显示出强烈的免疫标记GDNF在受损的脊髓运动神经元，反应性星形胶质细胞和少突胶质细胞，和表达病毒诱导的人GDNF mRNA转录在脊髓组织。Nissl染色细胞计数显示，注射AxCAhGDNF显著防止损伤的脊髓运动神经元在损伤后2-4周的丢失。这些结果表明，腺病毒介导的GDNF基因转移可以防止成年人脊髓损伤后运动神经元的变性。

项目成果

内田研造: "特集 脊柱靭帯骨化症.脊髄の可塑性と脊髄機能-圧迫下脊髄における生存活性と可塑性-"THE BONE. 16. 235-239 (2002)

Kenzo Uchida：“专题脊髓韧带骨化。脊髓可塑性和脊髓功能 - 受压下脊髓的生存活动和可塑性”THE BONE。

Uchida, K: "Progressive changes of neurofilament proteins and growth-associated protein-43 immunoreactivities at the site of cervical spinal cord compression in spinal hyperostotic mice"Spine. 27. 480-486 (2002)

Uchida, K：“脊柱骨质增生小鼠颈髓受压部位神经丝蛋白和生长相关蛋白 43 免疫反应性的进行性变化”Spine。

内田研造: "リポフェクチン法を用いた圧迫損傷脊髄に対する神経栄養因子の導入"厚生労働省特定疾患対策研究事業 脊柱靭帯骨化症に関する調査研究班 平成12年度研究報告書. 19-22 (2001)

Kenzo Uchida：“使用Lipofectin方法将神经营养因子引入受压脊髓”厚生劳动省特定疾病对策研究项目，脊柱韧带骨化研究小组，2000财年研究报告19-22（2001年）。

Uchida, K: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica. (in pressed). (2003)

Uchida, K：“脊柱骨质增生小鼠机械压缩的脊髓中神经营养素及其受体的表达增加（twy/twy）”《神经病理学报》。

Uchida, K., Baba, H., Furukawa, S., Omiya, M., Kokubo, Y., Kubota, C., Nakajima.: "Increased expression of neurotrophins and their receptors in the mechanically compressed spinal cord of the spinal hyperostotic mouse (twy/twy)"Acta neuropathologica.. (inp

Uchida, K.、Baba, H.、Furukawa, S.、Omiya, M.、Kokubo, Y.、Kubota, C.、Nakajima.：“脊髓机械压缩的脊髓中神经营养因子及其受体的表达增加