血管内皮生长因子转染胚胎成骨细胞的实验研究  

Transfection of embryonic osteoblasts by vascular endothelial growth factor： an experimental study

作　　者：于亚东[1] 刘增兵[1] 雷芳[1] 邵新中[1] 马涛[1] 毕伟东[1] 

机构地区：[1]河北医科大学附属第三医院河北省骨科医院手外科,石家庄050051

出　　处：《中华手外科杂志》2010年第4期239-242,共4页Chinese Journal of Hand Surgery

基　　金：河北省科技厅基金资助项目（052761197）

摘　　要：目的观察血管内皮生长因子（vascular endothelial growth factor，VEGF）转染胚胎成骨细胞后的成骨细胞增殖情况。方法取SD大鼠的胎鼠颅骨成骨细胞作原代培养后再进行体外培养扩增传代，同时构建含有VEGF基因片段（ad5-h-VEGF）的人5型腺病毒载体，以进入对数生长期的成骨细胞为靶细胞进行转染，免疫组化、免疫荧光定性检测实验组即转染（＋）组与对照组即转染（-）组细胞，绘制转染前后的生长曲线，并进行统计分析。结果VEGF在成骨细胞的表达可通过免疫组化、免疫荧光定性检测；实验组细胞增殖能力较对照组增强，统计分析转染组的细胞计数（〉48h）显著高于空白对照组（P〈0．05）。结论VEGF能够在成骨细胞中成功表达，VEGF转染后能够促进成骨细胞生长及成骨。Objective To investigate osteoblast proliferation of embryonic osteogenic cells transfeeted by vascular endothelial growth factor （VEGF）. Methods The osteoblasts isolated from fetal SD rat＇s cranium were cultured for primary and subsequent passages. Human type 5 adenovims construct carrying VEGF gene fragment （ad5-h-VEGF） was constructed and used to transfect osteoblasts that entered logarithm growth phase. Immanohistocbemistry and immunofluorescence staining of the cells with （transfectian group） or without （control group） transfection were carried out to measure expression of transfected genes in target cells and obtain growth curves. Statistical analysis was done. Results VEGF expression in the osteoblasts could be quantitatively detected by immunohistochemistry and immunofluorescence staining. Cells of the VEGF transfection group had stronger proliferation than cells of the control group. There were significantly more ostcoblasts in the trensfected group titan in the control group （ P 〈 0.05 ） after a certain time of growth （ 〉 48 h）. Conclusion Ad5-h- VEGF vector can successfully transfect osteoblasts and induce VEGF expression in osteoblasts. VEGF transfection promotes cell growth and bone formation.

关 键 词：成骨细胞 转染 细胞 培养的 血管内皮生长因子 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]