Tai Ming-Hong, Cheng Henrich, Wu Jen-Pey, Liu Yuh-Ling, Lin Pei-Ru, Kuo Jon-Son, Tseng Ching-Jiunn, Tzeng Shun-Fen
Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
Exp Neurol. 2003 Oct;183(2):508-15. doi: 10.1016/s0014-4886(03)00130-4.
Neuronal cell death and the failure of axonal regeneration cause a permanent functional deficit following spinal cord injury (SCI). Administration of recombinant glial cell line-derived neurotrophic factor (GDNF) has previously been reported to rescue neurons following severe SCI, resulting in improved hindlimb locomotion in rats. In this study, thus, GDNF gene therapy using an adenoviral vector (rAd-GDNF) was examined in rats following SCI induced by dropping the NYU weight-drop impactor from a height of 25 mm onto spinal segment T9-T10. To evaluate the efficacy of intraspinal injection of recombinant adenovirus into the injured spinal cord, we observed green fluorescent protein (GFP) gene transfer in the contused spinal cord. GFP was effectively expressed in the injured spinal cord, and the most prominently transduced cells were astrocytes. The expression of GDNF was detected only in rats receiving rAd-GDNF, not the controls, and remained detectable around the injured site for at least 8 days. Open-field locomotion analysis revealed that rats receiving rAd-GDNF exhibited improved locomotor function and hindlimb weight support compared to the control groups. Immunohistochemical examination for the neuronal marker, calcitonin gene-related peptide (CGRP), showed an increase in CGRP+ neuronal fibers in the injured spinal cord in rats receiving rAd-GDNF treatment. Collectively, the results suggest that adenoviral gene transfer of GDNF can preserve neuronal fibers and promote hindlimb locomotor recovery from spinal cord contusion. This research should provide information for developing a clinical strategy for GDNF gene therapy.
神经元细胞死亡和轴突再生失败会导致脊髓损伤（SCI）后出现永久性功能缺陷。先前有报道称，给予重组胶质细胞源性神经营养因子（GDNF）可在严重脊髓损伤后挽救神经元，从而改善大鼠的后肢运动能力。因此，在本研究中，我们对采用腺病毒载体（rAd-GDNF）进行GDNF基因治疗的大鼠进行了研究，这些大鼠的脊髓损伤是通过将纽约大学重物撞击器从25毫米高度 dropped 到脊髓节段T9-T10上诱导产生的。为了评估将重组腺病毒脊髓内注射到受损脊髓中的疗效，我们观察了挫伤脊髓中的绿色荧光蛋白（GFP）基因转移情况。GFP在受损脊髓中有效表达，转导最显著的细胞是星形胶质细胞。仅在接受rAd-GDNF的大鼠中检测到GDNF的表达，对照组未检测到，并且在损伤部位周围至少8天内均可检测到。旷场运动分析显示，与对照组相比，接受rAd-GDNF的大鼠运动功能和后肢负重能力有所改善。对神经元标志物降钙素基因相关肽（CGRP）进行的免疫组织化学检查显示，接受rAd-GDNF治疗的大鼠受损脊髓中CGRP+神经元纤维增加。总体而言，结果表明GDNF的腺病毒基因转移可以保留神经元纤维并促进脊髓挫伤后后肢运动功能的恢复。这项研究应为制定GDNF基因治疗的临床策略提供信息。
