IGF-1 gene transfer into denervated rat laryngeal muscle
IGF-1 gene transfer into denervated rat laryngeal muscle
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DOI：
10.1001/archotol.125.3.274
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发表时间：
1999-03-01
影响因子：
--
通讯作者：
O'Malley, BW
中科院分区：
文献类型：
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作者：
Flint, PW;Shiotani, A;O'Malley, BW
目的：利用报告基因、β -半乳糖苷酶和含有人IGF-1 （hIGF-1）基因序列的肌肉特异性表达系统，证明基因在大鼠喉肌中的转移，并研究hIGF-1基因在失神经支配的大鼠喉肌中的肌营养和神经营养作用。方法：将含有β -半乳糖苷酶DNA的聚乙烯基制剂注射到8只成年大鼠去神经甲状腺样肌中。12只动物同样服用了含有肌肉特异性表达系统和hIGF-1 DNA的聚乙烯基配方。12只动物注射等渗氯化钠溶液，均存活1个月。利用免疫组织化学技术检测β -半乳糖苷酶和hIGF-1的产生。采用胆碱酯酶或银染色和生长相关蛋白免疫染色（GAP-43）评估hIGF-1对运动终板和神经发芽的影响。结果：8只动物中有7只通过免疫染色、x -半乳糖组织化学染色或两者同时染色检测到β -半乳糖苷酶。在冷冻切片标本中，8只动物中有3只的hIGF-1免疫反应性呈阳性。在连续切片中，在3个hIGF-1阳性标本中的2个中，GAP-43定位于hIGF-1表达区域。用hIGF-1治疗的4只动物中有2只出现神经萌芽和运动终板接触增加。结论：使用含有肌肉特异性基因表达系统的聚乙烯基配方证明了基因转移到喉肌中。初步研究结果表明，在用hIGF-1载体处理的动物中，对运动终板、神经发芽和GAP-43的表达有积极作用。本研究为研究hIGF-1基因转移作为喉麻痹的新治疗方法奠定了基础。需要进一步的研究来量化肌营养和神经营养效应并确定治疗效益。
Objectives: To demonstrate gene transfer in rat laryngeal muscle using a reporter gene, beta-galactosidase, and a muscle-specific expression system containing the human IGF-1 (hIGF-1) gene sequence and to investigate the myotrophic and neurotrophic effects of hIGF-1 gene transfer in denervated rat laryngeal muscle.Methods: In 8 adult rats, a polyvinyl-based formulation containing beta-galactosidase DNA was injected into denervated thyroarytenoid muscle. Twelve animals were similarly administered a polyvinyl-based formulation containing a muscle-specific expression system and hIGF-1 DNA. Twelve animals were injected with isotonic sodium chloride solution, and all animals survived for 1 month. The production of beta-galactosidase and hIGF-1 was detected using immunohistochemical techniques. The effects of hIGF-1 on motor endplates and nerve sprouting were assessed using cholinesterase or silver staining and immunostaining for growth-associated protein (GAP-43).Results: beta-Galactosidase was detected in 7 of 8 animals by immunostaining, X-gal histochemical staining, or both. In frozen section specimens, hIGF-1 immunoreactivity was positive in 3 of 8 animals. In sequential sections, GAP-43 was localized to areas of hIGF-1 expression in 2 of the 3 hIGF-1-positive specimens. Increased nerve sprouting and motor endplate contact occurred in 2 of 4 animals treated with hIGF-1.Conclusions: Gene transfer into laryngeal muscle was demonstrated using a polyvinyl-based formulation containing a muscle-specific gene expression system. Preliminary findings indicate a positive effect on motor endplates, nerve sprouting, and the expression of GAP-43 in animals treated with the hIGF-1 vector. This study establishes a foundation for investigating hIGF-1 gene transfer as a novel treatment of laryngeal paralysis. Further studies are necessary to quantify myotrophic and neurotrophic effects and to establish therapeutic benefit.