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.
出处
《中华烧伤杂志》
CAS
CSCD
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 )
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