Heng Guan, Yongjun Li, Yuehong Zheng, Changwei Liu, Jing Yang, Cunxian Song, Pengyan Wang, Sanmei Zhao, Zongli Wang, Mingpeng She
Department of General Vascular Surgery, Peking Union Medical College Hospital, CAMS & PUMC, Beijing 100730.
Chin Med Sci J. 2002 Dec;17(4):220-4.
To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.
Nanoparticle-DNA complex was prepared with Poly-dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-1), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 microg), 6 with A-MCP-1 (200 microg) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed.
The package efficiency of the nanoparticle-DNA complex was 0.9%, release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300 nm. SMC genomic DNA PCR showed that A-MCP-1 gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-1 mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced.
Nanoparticle can act as a vector to transfect specific gene.
评估纳米颗粒作为新型载体用于特定基因转移的可能性及效率。
用携带反义单核细胞趋化蛋白-1(A-MCP-1,一种特异性表达基因)的聚-dl-乳酸-乙醇酸共聚物(PLGA)制备纳米颗粒-DNA复合物,测定其包裹效率、体外释放进程及复合物大小。以阳离子脂质体为对照,通过将基因转入培养的平滑肌细胞(SMC),采用基因组DNA聚合酶链反应(PCR)评估新型载体的可能性。在体内研究中,对20只新西兰白兔进行颈静脉-动脉搭桥移植手术,其中6个移植物用纳米颗粒-A-MCP-1(200微克)进行基因转移,6个用阳离子脂质体转染A-MCP-1(200微克),4个用LNCX质粒转染,4个作为对照。移植物取出14天后,通过斑点杂交检测静脉移植物中A-MCP-1的表达及其对MCP-1的影响,并对移植物进行形态学评估。
纳米颗粒-DNA复合物的包裹效率为0.9%,体外释放进程持续2周,大小范围为150至300纳米。SMC基因组DNA PCR显示纳米颗粒可成功将A-MCP-1基因转染入细胞。体内研究表明,纳米颗粒和脂质体局部基因递送组均有A-MCP-1 mRNA表达,同时静脉移植物中MCP-1表达显著受到抑制,新生内膜增生明显减少。
纳米颗粒可作为转染特定基因的载体。