CAJ | 학술논문

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(＞48 h)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.
목적 관찰혈관내피생장인자(vascular endothelial growth factor,VEGF)전염배태성골세포후적성골세포증식정황.방법 취SD대서적태서로골성골세포작원대배양후재진행체외배양확증전대,동시구건함유VEGF기인편단(ad5-h-VEGF)적인5형선병독재체,이진입대수생장기적성골세포위파세포진행전염,면역조화、면역형광정성검측실험조즉전염(+)조여대조조즉전염(-)조세포,회제전염전후적생장곡선,병진행통계분석.결과 VEGF재성골세포적표체가통과면역조화、면역형광정성검측;실험조세포증식능력교대조조증강,통계분석전염조적세포계수(＞48 h)현저고우공백대조조(P＜0.05).결론 VEGF능구재성골세포중성공표체,VEGF전염후능구촉진성골세포생장급성골.
Objective To investigate osteoblast proliferation of embryonic osteogenic cells transfected 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 adenovirus construct carrying VEGF gene fragment (ad5-h-VEGF) was constructed and used to transfect osteoblasts that entered logarithm growth phase.Immunohistochemistry and immunofluorescence staining of the cells with (transfection group) or without ( control group) transfecfion 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 osteoblasts in the trensfected group than in the control group ( P ＜ 0. 05) after a certain time of growth ( ＞ 48 h). Conclusion Ad5-hVEGF vector can successfully transfect osteoblasts and induce VEGF expression in osteoblasts. VEGF transfection promotes cell growth and bone formation.