摘要
目的:构建pcDNA3.1/VEGF165质粒及联合脱蛋白骨基因治疗早期股骨头缺血性坏死。方法:将含人VEGF165全长cDNA序列的克隆载体pUC18/VEGF165与pcDNA3.1(+)载体构建成重组真核表达质粒pcDNA3.1/VEGF165;69只AVNFH造模成功后的新西兰大白兔随机分成3组。第一组,将脱蛋白骨复合pcDNA3.1/VEGF165质粒植入坏死的股骨头内;第二组植入DPB;第三组仅在股骨头内钻一隧道。股骨头标本术后3天,1,2,4,8和16周获取。RT-PCR检测VEGF165mRNA的表达,Western blot检测VEGF165蛋白的表达,组织形态学分析血管发生和股骨头修复。结果:成功构建pcDNA3.1/VEGF165;第一组VEGF165 mRNA和蛋白表达术后1周达到高峰,时间持续超过3周。血管形成术后2,4周增加,骨形成术后2,4和8周增加,与另两组相比差异呈显著性(P<0.01)。结论:VEGF165基因转染可促进局部血管的早期形成,DPB-VEGF复合物可增加骨形成。脱蛋白骨联合VEGF165基因治疗为骨坏死的修复提供了理论基础。
Objective：To reconstruct pcI）NA.3.1/VEGF165 and combine it with deproteinized bone（DPB） to repair avascular necrosis of the femoral head （AVNFH）. Methods ：Clone plasmid pUC18/VEGF165 containing the whole cDNA sequence of VEGF165 was combined with pcDNA.3.1 （ ＋ ） to reconstruct pcDNA3.1/VEGF165. Sixty-nine AVNFH model New Zealand adult rabbits with a mean weight of 2.8 kg were randomly divided into three groups. In group 1 ,DPB combined with the reconstructed plasmid pcDNA3.1/VEGF165 was implanted in the drilled channel of the necrotic femoral head. In group 2 ,only DPB was implanted. In group 3 ,only channel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained 3 days, 1,2,4, and 8 weeks after operation. The expression of vascular endothelial growth factor （VEGF） was examined by RT- PCR, Western blotting techniques. Angiogenesis and repair of the femoral head were observed by histological and histomorphometric analysis. Results ： pcDNAB, 1/VEGF165 was reconstructed successfully. [n group 1, the expression of VEGF was detected in the femoral head implanted by DPB - VEGF compound. VEGF165 mRNA and protein detected went up to apex 1 week postoperatively and lasted more than 3 weeks. The femoral head implanted by DPB- VEGF compound showed a significant difference in vascularization 2 and 4 weeks after operation and a significant difference in bone formation 2,4 and 8 weeks after operation from those in other groups on histomorphometric analysis （P〈0.01 ）. Conclusion ：Transfection of VEGF165 gene enhances local angiogenesis in early time and DPB- VEGF compound improves bone formation. Deproteinized Done combined with VEGF gene provides a potential method for treatment of osteonecrcxsis.
出处
《重庆医科大学学报》
CAS
CSCD
2005年第4期505-508,547,共5页
Journal of Chongqing Medical University
基金
重庆市卫生局项目(编号:002010)
关键词
脱蛋白骨
血管内皮生长因子
基因治疗
股骨头缺血坏死
Deproteinized bone
Vascular endothelial growth factor
Gene therapy
Avascular necrosis of the femoral head
