Department of Orthopaedics and Rehabilitation Medicine, The University of Fukui, Fukui, Japan.
Spine (Phila Pa 1976). 2010 Mar 1;35(5):497-504. doi: 10.1097/BRS.0b013e3181b8e89b.
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.
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.
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.
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.
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.
Our results suggest that targeted retrograde BDNF gene delivery suppresses apoptosis of neurons and oligodendrocytes in the injured rat spinal cord.
通过肌肉内注射靶向逆行腺病毒（AdV）介导的脑源性神经营养因子（BDNF）基因转染治疗脊髓损伤大鼠的组织学和免疫组织化学研究。
探讨靶向逆行 AdV-BDNF 基因转染对创伤性脊髓损伤的神经保护作用，特别是在预防神经元和少突胶质细胞凋亡方面的作用。
多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，以预防损伤脊髓中神经细胞凋亡。然而，尚无报道描述靶向逆行神经营养因子基因转染对预防神经细胞凋亡在损伤脊髓中的潜在作用。
在大鼠脊髓损伤后立即通过双侧胸锁乳突肌逆行给予 AdV-BDNF 或 AdV-LacZ，以进行逆行传递。通过免疫荧光染色和细胞标志物（NeuN、RIP、GFAP、OX-42 和 NG2）的双重染色，检测 LacZ 基因或 AdV-BDNF 基因转染产生的β-半乳糖苷酶表达的定位。基因注射后，计数 TUNEL 阳性细胞并检测活性 caspase-3 和 NG2 的免疫反应性。
LacZ 标记基因的逆行传递在颈段脊髓神经元和包括白质中的少突胶质细胞在内的神经胶质细胞中被识别。与 AdV-LacZ 注射相比，AdV-BDNF 转染通过下调半胱天冬酶凋亡途径，显著减少 TUNEL 阳性凋亡细胞的数量，并显著促进损伤脊髓中 NG2 的表达。
我们的结果表明，靶向逆行 BDNF 基因转染可抑制损伤大鼠脊髓中神经元和少突胶质细胞的凋亡。
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