熊蠡茗;郭兵;邵增务;杨述华;谢卯;王河忠;

• 1:华中科技大学同济医学院附属协和医院骨科

摘要(Abstract)：

[目的]考察腺病毒载体搭载的重组人组织金属蛋白酶抑制因子-3基因(RAdTIMP-3)转染兔椎间盘后对椎间盘内主要基质成分质量的影响,并探讨其用于椎间盘退变治疗的可行性。[方法]扩增Lac-Z基因标记的RAdTIMP-3和重组腺病毒载体RAd66,并进行纯化、鉴定及滴度测定。将30只日本大白兔随机分入5组,分别向L4、5、L5、6椎间盘内注射生理盐水(第1组)、1.0×1010OPU/mlRAd66(第2组)以及1.0×1010OPU/mlRAdTIMP-3(第3、4、5组)各25μl。各组分别于注射后2、2、1、2、4周收集标本,X-gal染色检测转染效率,RT-PCR检测TIMP-3及聚集蛋白聚糖核心蛋白的表达,TIMP-3及H型胶原免疫组化染色、藏红O-快绿染色及Tunel染色考察转染对椎间盘基质的调控作用。[结果](1)RAdTIMP-3经扩增和纯化后浓度可达1.9×10TM OPU/ml。(2)RT-PCR显示,3、4、5组TIMP-3基因表达均较1、2组明显增加,3、4、5组核心蛋白基因表达较1、2组略有增强。(3)Tunel染色显示各组凋亡细胞比例无明显差异。(4)第5组藏红O-快绿染色及II型胶原免疫组化染色染色强度均好于1、2组。[结论]RAdTIMP-3能够广泛而安全地在兔椎间盘内表达,并改善椎间盘基质成分的质和量,具备用于椎间盘退变治疗的潜力。

关键词(KeyWords)： 组织金属蛋白酶抑制因子-3;椎间盘;聚集蛋白聚糖;Ⅱ型胶原

Abstract:

Keywords:

基金项目(Foundation): a grant from the National Natural Sciences Foundation of China(No:30700841)

作者(Author): 熊蠡茗;郭兵;邵增务;杨述华;谢卯;王河忠;

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DOI:

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