人骨形态发生蛋白2重组腺病毒在动物模型中活性观测

Study of the Human Bone Morphogenetic Protein-2 Recombinant Adenovirus in the Treatment of Bone Defect

目的 利用基因工程技术构建人骨形态发生蛋白 2重组腺病毒 ,并进行体内表达 ,测定活性 ,为深入开展的基因治疗研究创造条件。方法 同源重组构建重组腺病毒 ,将 30只SD大鼠随机分为治疗组和对照组 ,将纯化后的重组腺病毒 5 0 μl注入SD大鼠股部肌肉陷窝病损处 ,通过组织学染色、X线片对动物模型的组织变化进行观测。结果 重组腺病毒治疗组有明显的诱导成骨活性 ,治疗组碱性磷酸酶活性明显高于对照组。结论 应用于动物实验中目的基因表达 ,并具有生物学活性。本研究是在骨缺损基因治疗方面的一次尝试 ,且人骨形态发生蛋白 2重组腺病毒的构建成功 ,也为我国骨科疾病的基因治疗进一步研究奠定了基础。

Objective To construct the recombinant adenovirus of human bone morphogenetic protein-2 and to detect its expression and activity in tissues. Methods The cDNA coding for the full length BMP2 was amplified, purified, and inserted into the vector PGEM-T-Easy. The recombinant virus was constructed using the DNA/calcium phosphate precipitate containing PGEM-T-BMP2 and PJM17, 30 SD rats were divided into two groups randomly and 50 μl purified recombinant adenovirus was injected into the calf muscle of the rats.The expression of BMP gene in muscle tissue was measured and the changes of ALP staining and X-ray examination were compared. Results Significant activity of bone induction was found in the treatment group with the recombinant adenovirus of hBMP2, compared with the normal control group. The level of ALP activity in the muscular tissues of the treatment group was higher than that in the control group. Conclusion The recombinant AdBMP2 was constructed and the hBMP2 is functionally active as demonstrated by the induction of ALP activity in the muscular tissues of animal model.