摘要
目的 观察纳米粒子携带血管内皮生长因子（VEGFl65）基因转染的可行性。方法 应用聚乳酸聚乙醇酸共聚物（PL-GA）和聚乙烯醇（PVA）包载VEGFl65基因质粒，制备纳米级粒子混合物，检测其载药量、体外释放情况及粒径；培养心肌细胞，应用RT-PCR及ELISA方法观察纳米级粒子混合物为真核细胞传递基因的可行性；将纳米粒子悬浮液注射至活体家兔心肌组织内，96h后电镜观察其向心肌组织内传递基因的可行性。结果 制备的纳米粒子载药量为1．87％，粒径为25～300nm；RT-PCR和ELISA结果提示纳米粒子可将目的基因转移至心肌细胞中；体内实验显示心肌细胞胞质内及胞核内可见大量被吞噬的纳米粒子。结论 纳米粒子可作为向心肌组织转运VEGF基因的载体。
Objective To evaluate the possibility and efficiency of nanoparticles as a new vector in vascular endothelial growth factor （VEGF） gene transfection. Methods Nanoparticle-VEGF （Np/VEGF）complex was prepared with poly （D, L-lactide co-glycolide） （PLGA） loading VEGF165 gene using the multiple emulsion （w/o/w） technique. The envelopment efficiency and size of the complex were determined. Rat myocardial cells were cultured in vitro, and the Np/VEGF was transfected into the cultured myocardial cells. Then RT-PCR and ELISA were used to evaluate whether the Np/VEGF increased the level of gene expression. Four New Zealand rabbits were used, the suspension of Np/VEGF was injected into myocardial tissue of rabbits after thoracotomy. 96h after the operation, the tissue sections of the implant sites were observed with transmission electron microscope （TEM） to determine the process of nanoparticles as vectors for gene transfer to cardiac myocytes. Results The envelopment efficiency and size of the Np/VEGF complex thus prepared were 1.87% and 25-300nm respectively. RT-PCR and ELISA showed that VEGF gene could be successfully transfected into myocardial cells by nanoparticle, and NP/VEGF significantly enhanced gene transfection efficiency, and it was more effective than plasmid. 96h after the operation, a great number of nanoparticles were observed in myocardial cytoplasm and nucleus with TEM, and many nanoparticles began to dissolve and degrade, suggesting that the DNA was released slowly from the nanoparticles localized in the cytoplasmic compartment, and was then transferred into the nucleus. Condusions NP/VEGF can act as a vector to transfect VEGF gene in vitro and in vivo, it significantly enhanced gene transfection efficiency, and it was more effective than plasmid.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2006年第3期213-215,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金(30300456)
关键词
转染
纳米粒子
血管内皮生长因子类
transfection
nanoparticle
vascular endothelial growth factors
