血管内皮细胞生长因子165基因转染对兔骨缺损修复的影响  

Effect of vascular endothelial growth factor 165 gene transfection on repair of bone defect:experiment with rats

作　　者：赵冬梅[1] 杨加峰[2] 武士清[1] 邱丽萍[2] 刘军莉[3] 王海斌[1] 许复郁[4] 蔡景龙[2] 

机构地区：[1]山东大学第二附属医院骨科,济南250033 [2]山东大学第二附属医院整形外科 [3]山东大学第二医院分子实验室 [4]山东大学第二医院病理科

出　　处：《中华医学杂志》2007年第25期1778-1782,共5页National Medical Journal of China

基　　金：山东省科技攻关计划资助项目(032050112);山东省自然科学基金(Q2006C08);山东省卫生厅科研 计划(2003)

摘　　要：目的构建 pcDNA3.1-血管内皮细胞生长因子(VEGF)165质粒,观察其对兔骨缺损修复的影响。方法构建成重组真核表达质粒 pcDNA3.1-VEGF165;采用30只新西兰大白兔,制备双侧桡骨骨缺损模型,实验组以体积等大的明胶海绵置于创腔,局部直接注射稀释的质粒液200 ng;对照组以同量生理盐水代替质粒液,同法处理。于术后1、2、4、6、8和12周行 X 线照片后获取标本,观察缺损局部组织修复及新生血管并计算微血管密度(MVD)。结果 pcDNA3.1-VEGF165质粒构建成功,测序正确。术后 X 线:1周时两组无明显差别,实验组2周骨痂形成、骨质修复,12周时骨质已正常,对照组表现为修复迟缓;HE 染色:实验组2周大量新生血管形成、通血,4周成骨细胞大量增殖、骨小梁形成,12周骨皮质改建完成,骨髓腔再通;对照组经历过程类似,但相对时间延迟,12周时骨髓腔及皮质改建尚未完成。2周时对照组及实验组 MVD 分别为56.1±6.1、69.1±5.4,均较1周时42.2±6.4、47.0±7.5时增加,组间及组内间差异有统计学意义(t=8.0347,P=0.0000)。结论骨缺损局部直接应用 PcDNA3.1-VEGF165质粒液后,可表达并上调 VEGF165,具有促进局部微血管形成、加快骨缺损修复的作用。Objective To investigate the influence of vascular endothelial growth factor （VEGF） 165 gene transfection on the repair of bone defect. Methods 38 New Zealand rabbits underwent resection of a segment 1 cm in length in bilateral radii filled with absorbable gelatin sponge. Dilated solution of the plasmid pcDNA3.1/ VEGF165 was injected into the bone defect of one side and normal saline was injected into the contralateral bone defect. 1, 2, 4, 6, 8, and 12 weeks later X ray examination was conducted to observe the repair of bone defect, and then 5 rabbits were killed at each time points to take out the bone defects. HE staining was used to observe the bone repair. The levels of mcrovessel density （MVD） 1 and 2 weeks after the operation were observed. RT-PCR was used to detect the mRNA expression of VEGF in the bone defect. Based on the results of RT-PCR the tissue mRNA expression of VEGF65 was detected by realtime quantitative polymerase chain reaction （RQ-PCR）. Results X-ray examination showed that there was no significant difference in the wound healing between the two group 1 week after the operation in all rabbits. Some callus could be seen in the experimental group 2 weeks after. Twelve weeks after the operation the reconstruction of bone cortex was completed. Similar process occurred in the control sides but more lately. The MVD level 7 days after of the experimental group was 47.0 ±7.5, significantly higher than that of the control group （42.2 ±6.4, t =2. 4519, P = 0. 0179）, and the MVD level 14 days after of the experiment group was 69.1± 5.4, significantly higher than that of the control group （ 56. 1 ± 6.1, t = 8. 0347, P = 0. 0000）. In the experimental group the mRNA expression amounts of VEGF165 could be found 1 week after, gradually increased and peaked 3 weeks after, then decreased, and became stable 6 weeks after. The mRNA expression amounts of VEGF165 in the control group were lower than those of the experimental group.Conclusion Local application of PcDNA3. 1/VEGF165 vector p

关 键 词：基因转移 垂直 内皮 血管 内皮细胞生长因子 

分 类 号：R68[医药卫生—骨科学]