Timing of human insulin-like growth factor-1 gene transfer in reinnervating laryngeal muscle
Timing of human insulin-like growth factor-1 gene transfer in reinnervating laryngeal muscle
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DOI：
10.1097/00005537-200404000-00024
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发表时间：
2004-04-01
期刊：
影响因子：
2.6
通讯作者：
Flint, PW
中科院分区：
文献类型：
--
作者：
Nakagawa, H;Shiotani, A;Flint, PW
目的/假设：作者设计了一种大鼠喉麻痹模型，以研究使用肌肉特异性表达系统的基因转移策略，以增强人胰岛素样生长因子-1（hIGF-1）的局部递送。在初步研究中，含有α-肌动蛋白启动子和人hIGF-1序列的非病毒载体产生神经营养和肌营养作用1个月后，单次注射质粒制剂到瘫痪的大鼠甲杓肌在体内。基于这些发现，假设hIGF-1的作用将增强喉肌神经支配程序的结果。基因递送相对于神经修复的时机可能很重要，以优化结果。研究设计：前瞻性分析。研究方法：在喉返神经切断和修复后立即治疗的大鼠和神经切断和修复后30天接受延迟治疗的大鼠中评价了非病毒基因转移对hIGF-1递送的影响。采用聚合酶链反应和逆转录-聚合酶链反应技术测定基因转移效率。检查肌纤维直径、运动终板长度和与神经接触的运动终板的百分比以评估hIGF-1的营养作用。结果如下：与再神经化未处理的对照样品相比，早期和延迟的hIGF-1转移导致肌纤维直径的显着增加。运动终板长度显着减少，神经/运动终板接触显着增加后，延迟基因转移，但不是在早期治疗。结论：我们从研究结果中推断，通过单次肌肉注射延迟hIGF-1基因转移将增强肌肉神经再支配的过程。这些研究结果的临床意义支持了未来应用非病毒载体基因治疗喉麻痹和其他周围神经损伤的管理。
Objectives/Hypothesis: The authors have designed a rat laryngeal paralysis model to study gene transfer strategies using a muscle-specific expression system to enhance local delivery of human insulin-like growth factor-1 (hIGF-1). In preliminary studies, a nonviral vector containing the alpha-actin promoter and human hIGF-1 sequence produced both neurotrophic and myotrophic effects 1 month after single injection of plasmid formulation into paralyzed rat thyroarytenoid muscle in vivo. Based on these findings, it is hypothesized that the effects of hIGF-1 will enhance the results of laryngeal muscle innervation procedures. The timing of gene delivery relative to nerve repair is likely to be important, to optimize the results. Study Design: Prospective analysis. Methods: The effects of nonviral gene transfer for the delivery of hIGF-1 were evaluated in rats treated immediately following recurrent laryngeal nerve transection and repair and in rats receiving a delayed treatment schedule, 30 days after nerve transection and repair. Gene transfer efficiency was determined using polymerase chain reaction and reverse transcriptase-polymerase chain reaction techniques. Muscle fiber diameter, motor endplate length, and percentage of motor endplates with nerve contact were examined to assess hIGF-1 trophic effects. Results: Compared with reinnervated untreated control samples, both early and delayed hIGF-1 transfer resulted in significant increase in muscle fiber diameter. Motor endplate length was significantly decreased and nerve/motor endplate contact was significantly increased following delayed gene transfer, but not after early treatment. Conclusion: We infer from results of the study that delayed hIGF-1 gene transfer delivered by a single intramuscular injection will enhance the process of muscle reinnervation. The clinical relevance of these findings supports the future application of gene therapy using nonviral vectors for management of laryngeal paralysis and other peripheral nerve injuries.