圈套寡核苷酸抑制NIH3T3细胞α2Ⅰ型胶原基因表达的研究  

An experimental study of inhibiting effect of decoy oligodeoxynucleotides on the gene expression of collagen in NIH3T3 cells

作　　者：郝天智[1] 梁华平[1] 吕凤林[1] 徐祥[1] 国华[2] 王付龙 杨文军[4] 罗艳[1] 李磊[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所 [2]泰安解放军第八十八医院 [3]邯郸解放军第二五八医院 [4]重庆医科大学血管外科

出　　处：《中华烧伤杂志》2003年第3期175-178,共4页Chinese Journal of Burns

基　　金：国家自然科学基金资助项目 (C0 3 0 2 0 4) ;国家重点基础研究发展规划资助项目 (G19990 5 42 0 3 ) ;重庆市卫生局课题

摘　　要：目的　研究激活剂蛋白 1(activatorprotein 1,AP 1)圈套寡核苷酸 (decoy oligodeoxynu cleotides,Decoy ODNs)对成纤维细胞α2Ⅰ型胶原表达的影响 ,探讨病理性瘢痕的基因治疗。　方法　设计合成针对AP 1的Decoy ODNs,用阳离子脂质体转染NIH3T3细胞 ,观察Decoy ODNs在细胞中的分布 ;用凝胶迁移变动分析 (electrophoreticmobilityshiftassay ,EMSA)研究Decoy ODNs对AP 1的抑制作用 ,采用RT PCR观察其对细胞胶原合成的影响。　结果　AP 1的Decoy ODNs可在体外竞争抑制核转录因子AP 1的活性 ;阳离子脂质体可以将Decoy ODNs转染进入细胞浆及细胞核从而发挥作用 ;Decoy ODNs作用 2 4h后 ,NIH3T3细胞α2Ⅰ型胶原mRNA的表达明显降低。　结论　Decoy ODNs可以通过拮抗核转录因子AP 1的活性而抑制α2Ⅰ型胶的表达。Objective To investigate the effect of activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) on the expression of fibroblast α2 type I collagen, so as to explore the gene therapy of pathologic scar. Methods Decoy-ODNs targeting AP-1 were designed and synthesized. NIH3T3 cells were transfected by cationic liposomes. The distribution of Decoy-ODNs in the cells was investigated. The inhibiting effects of Decoy-ODNs on AP-1 were determined by electrophoretic mobility shift assay (EMSA). And the effects of Decoy-ODNs on the collagen synthesis in the cells were analyzed by RT-PCR. Results AP-1 Decoy-ODNs could competitively inhibit the AP-1 in vitro activity . Cationic liposomes could play roles by effectively transfecting Decoy-ODNs into the plasma and nucleus. The mRNA expression of fibroblast α2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action. Coclution Decoy-ODNs could inhibit the mRNA expression of fibroblast α2 type I collagen by antagonizing AP-1.

关 键 词：圈套寡核苷酸 NIH3T3细胞 α2I型胶原 基因表达 成纤维细胞 

分 类 号：R622[医药卫生—整形外科]