小鼠双微体扩增基因反义寡核苷酸在血管平滑肌细胞中的摄取动力学研究  

Research of mdm2 antisense oligonucleotide of the uptake kinetics in vascular smooth muscle cells

作　　者：付鹏[1] 韩巍[1] 姜廷军[1] 赵长久[1] 尹雪松[2] 

机构地区：[1]哈尔滨医科大学附属第四医院核医学科,黑龙江哈尔滨150001 [2]哈尔滨医科大学附属第四医院急诊科,黑龙江哈尔滨150001

出　　处：《哈尔滨医科大学学报》2011年第5期405-408,共4页Journal of Harbin Medical University

基　　金：黑龙江省教育厅科学技术研究项目(11531192)

摘　　要：目的研究小鼠双微体扩增基因(mouse double minute 2,mdm2)反义寡核苷酸(antisense oli-gonucleotide,ASON)在血管平滑肌细胞中的摄取动力学,探讨mdm2反义寡核苷酸包埋支架防治支架内再狭窄的可行性。方法人工合成一段针对mdm2 mRNA的反义寡核苷酸,以肼基尼古酰胺(HYNIC)作为双功能螯合剂进行99 Tcm标记,以脂质体包裹99 Tcm标记的ASON,不同时间检测脂质体包裹的ASON在血管平滑肌细胞中的摄取和清除情况,并与未经脂质体包裹的ASON在平滑肌细胞中的摄取及清除情况比较。结果 ASON的标记率为(52.30±2.03)%,经纯化后放化纯达95%以上。平滑肌细胞对脂质体包裹的ASON的摄取峰值为(48.22±2.39)%,且明显高于未经脂质体包裹的ASON,脂质体包裹的ASON的清除率在各时间点均低于未经脂质体包裹的ASON。结论 mdm2的反义寡核苷酸经脂质体包裹后,平滑肌细胞对反义寡核苷酸能达到较大的摄取比率,清除较缓慢。mdm2反义寡核苷酸有望被进一步应用于药洗脱支架研究。Objective To investigate the possibility of using mdm2 mRNA antisense oligonucleotide（ASON）on preventing restenosis following angioplasty,the uptake kinetics of the mdm2 ASON in vascular smooth muscle cells.Methods An ASON targeted to mdm2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator HYNIC.The uptake kinetics of the liposome-coated antisense oligonucleotide were studied in vascular smooth muscle cells at different times,compared with those of the non-coated ASON in vascular smooth muscle cells.Results The labeling efficiency of ASON was（52.30±2.03）% with the bifunctional chelator HYNIC.The radiochemical purity was above 95% after purification.The uptake peak value of the liposome-coated ASON was（48.22±2.39）%.The liposome-coated ASON showed significantly higher accumulation and effluxed much slowly than non-coated ASON.Conclusion The higher uptake rate and lower clearance rate of the liposome-coated mdm2 ASON can be achieved in vascular smooth muscle cells.The ASON can accumulate in vascular smooth muscle cells more specifically which provids a basis for it to be used in drug eluting stent study.

关 键 词：小鼠双微体扩增基因 平滑肌细胞 寡核苷酸 脂质体 

分 类 号：R817.5[医药卫生—影像医学与核医学]