脂质体介导VEGF基因对成骨细胞增殖、合成骨钙素以及细胞周期的相关研究  

Effect of Liposome Mediated VEGF Gene on Osteoblast Proliferation,Osteocalcin Synthesis and Cell Cycle

作　　者：熊军[1] 刘韦 曾凡[1] 陈剑飞[1] 周海燕[2] XIONG Jun;LIU Wei;ZENG Fan;CHEN Jian-fei;ZHOU Hai-yan(Department of orthopedics,Hainan Provincial People＇s Hospital,Haikou,Hainan,570311,China;Department of medicine,Affiliated Hospital ofHainan Meducal University Haikou,Hainan,570120,China)

机构地区：[1]海南省人民医院创伤骨科,海南海口570311 [2]海南医学院附属医院内科,海南海口570120

出　　处：《现代生物医学进展》2018年第16期3042-3045,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81260275)

摘　　要：目的:研究脂质体介导血管内皮生长因子(VEGF)基因对成骨细胞增殖、合成骨钙素以及细胞周期的影响。方法:通过脂质体介导的基因转染方法,将携带外源性VEGF重组pcDNA3-hVEGF质粒导入体外培养的成骨细胞,酶联免疫吸附测定法(ELISA)检测转染后细胞中VEGF浓度变化,以判断转染效果;采用细胞计数法检测转染重组质粒的成骨细胞的增殖活性;流式细胞术检测转染重组质粒的成骨细胞周期的变化;ELISA检测转染重组质粒的成骨细胞骨钙素浓度变化。结果:与对照组相比,转染组成骨细胞中VEGF的浓度显著增加,与对照组间差异具有统计学意义(P<0.05);转染重组质粒的成骨细胞的增殖能力较对照组显著增强,差异具有统计学意义(P<0.05),与对照组相比,转染重组质粒的成骨细胞周期(G2/M+S)%明显增加,差异具有统计学意义(P<0.05);转染重组质粒的成骨细胞合成的骨钙素浓度较对照组显著升高,差异具有统计学意义(P<0.05)。结论:脂质体介导成骨细胞增加血管内皮生长因子的水平,可促进成骨细胞增殖,增加成骨细胞骨钙素的浓度,从而提高成骨细胞的功能。Objective： To investigate the effects of vascular endothelial growth factor（VEGF） gene mediated by liposome on osteoblast proliferation,osteocalcin synthesis and cell cycle. Methods： The recombinant plasmid pcDNA3-hVEGF with exogenous VEGF was introduced into cultured osteoblasts by liposome-mediated gene transfection method. The concentration changes of VEGF were detected by enzyme linked immunosorbent assay（ELISA） in the transfected cells to determine transfection efficiency. The osteoblast proliferation were measured by Cell counting; the cell cycle were analyzed by flow cytometry; the expression of osteocalcin were detected by ELISA. Results： Compared with the control group,the concentration of VEGF in transfected osteoblasts increased significantly,and the difference was statistically significant（P〈0.05）. The proliferation of osteoblasts transfected with recombinant genes was significantly enhanced（P〈0.05）,Cell cycle（G2/M＋S）% increased significantly,and the difference was statistically significant（P〈0.05）; the concentration of synthetic osteocalcin was significantly increased,and the difference was statistically significant（P〈0.05）.Conclusion： Liposome-mediated osteoblasts increase vascular endothelial growth factor level,promote osteoblast proliferation and increase osteoblast osteocalcin concentration to improve osteoblast function.

关 键 词：VEGF 成骨细胞 增殖 骨钙素 

分 类 号：R-33[医药卫生] R68