Targeted Retrograde Gene Delivery of Brain-Derived Neurotrophic Factor Suppresses Apoptosis of Neurons and Oligodendroglia After Spinal Cord Injury in Rats
Targeted Retrograde Gene Delivery of Brain-Derived Neurotrophic Factor Suppresses Apoptosis of Neurons and Oligodendroglia After Spinal Cord Injury in Rats
复制标题
DOI：
10.1097/brs.0b013e3181b8e89b
复制
发表时间：
2010-03-01
期刊：
影响因子：
3
通讯作者：
Baba, Hisatoshi
中科院分区：
文献类型：
--
作者：
Nakajima, Hideaki;Uchida, Kenzo;Baba, Hisatoshi
研究设计.靶向逆行腺病毒（AdV）介导的脑源性神经营养因子（BDNF）基因通过肌肉注射在脊髓损伤大鼠中的组织学和免疫组织化学研究。目的：探讨AdV-BDNF基因在脊髓损伤后的神经保护作用，探讨AdV-BDNF基因在脊髓损伤后对神经元和少突胶质细胞凋亡的保护作用。几项研究调查了神经营养因子包括BDNF对脊髓损伤的神经保护作用，关于预防损伤脊髓中的神经细胞凋亡。然而，目前还没有关于靶向逆行神经营养因子基因导入损伤脊髓预防神经细胞凋亡的报道。在大鼠脊髓损伤后立即将AdV-BDNF或AdV-LacZ经双侧胸锁乳突肌逆行递送至脊髓副运动神经元。通过免疫荧光染色和细胞标记物（NeuN、RIP、GFAP、OX-42和NG 2）的双重染色来检查LacZ基因或AdV-BDNF基因转染产生的β-半乳糖苷酶表达在损伤脊髓中的定位。注射基因后，计数TUNEL阳性细胞，检测caspase-3和NG 2的免疫反应性。LacZ标记基因在脊髓神经元和神经胶质细胞（包括白色内的少突胶质细胞）中逆行表达;与AdV-LacZ相比，AdV-BDNF转染可通过下调caspase凋亡途径减少TUNEL阳性凋亡细胞数量，并显著促进NG 2表达。我们的研究结果表明，靶向逆行BDNF基因传递抑制损伤大鼠脊髓神经元和少突胶质细胞的凋亡。
Study Design. Histologic and immunohistochemical studies after targeted retrograde adenovirus (AdV)-mediated brain-derived neurotrophic factor (BDNF) gene delivery via intramuscular injection in rats with injured spinal cord.Objective. To investigate the neuroprotective effect of targeted retrograde AdV-BDNF gene transfection in the traumatically injured spinal cord in terms of prevention of apoptosis of neurons and oligodendrocytes.Summary of Background Data. Several studies investigated the neuroprotective effects of neurotrophins including BDNF on spinal cord injury, with respect to prevention of neural cell apoptosis in injured spinal cord. However, no report has described the potential effect of targeted retrograde neurotrophic factor gene delivery in injured spinal cord on prevention of neural cell apoptosis.Methods. AdV-BDNF or AdV-LacZ was used for retrograde delivery via bilateral sternomastoid muscles to the spinal accessory motoneurons immediately after spinal cord injury in rats. Localization of beta-galactosidase expression produced by LacZ gene or AdV-BDNF gene transfection was examined by immunofluorescence staining and double staining of cell markers (NeuN, RIP, GFAP, OX-42, and NG2) in the injured spinal cord. TUNEL-positive cells were counted and immunoreactivity to active caspase-3 and NG2 was examined after gene injection.Results. Retrograde delivery of LacZ marker gene was identified in cervical spinal neurons and glial cells including oligodendrocytes in the white matter.AdV-BDNF transfection resulted in a significant decrease in the number of TUNEL-positive apoptotic cells by downregulating the caspase apoptotic pathway, with significant promotion of NG2 expression in injured spinal cord, compared with AdV-LacZ injection.Conclusion. Our results suggest that targeted retrograde BDNF gene delivery suppresses apoptosis of neurons and oligodendrocytes in the injured rat spinal cord.