Hao Tian-zhi, Liang Huan-ping, Lv Feng-lin, Xu Xiang, Guo Hua, Wang Fu-long, Yang Wen-jun, Luo Yan, Li Lei
Department of neurosurgery, Research Institute of surgery, Daping Hospital, Third military Medical University, Chongqing 400042, PR China.
Zhonghua Shao Shang Za Zhi. 2003 Jun;19(3):175-8.
To investigate the effect of activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) on the expression of fibroblast alpha2 type I collagen, so as to explore the gene therapy of pathologic scar.
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.
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 alpha2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action.
Decoy-ODNs could inhibit the mRNA expression of fibroblast alpha2 type I collagen by antagonizing AP-1.
研究活化蛋白-1(AP-1)诱饵寡脱氧核苷酸(Decoy-ODNs)对成纤维细胞α2Ⅰ型胶原表达的影响,以探讨病理性瘢痕的基因治疗方法。
设计并合成针对AP-1的Decoy-ODNs。采用阳离子脂质体转染NIH3T3细胞,研究Decoy-ODNs在细胞内的分布情况。通过电泳迁移率变动分析(EMSA)检测Decoy-ODNs对AP-1的抑制作用。运用逆转录聚合酶链反应(RT-PCR)分析Decoy-ODNs对细胞内胶原合成的影响。
AP-1 Decoy-ODNs能够在体外竞争性抑制AP-1的活性。阳离子脂质体可通过将Decoy-ODNs有效转染至细胞质及细胞核发挥作用。Decoy-ODNs作用24小时后,成纤维细胞α2Ⅰ型胶原的mRNA表达明显降低。
Decoy-ODNs可通过拮抗AP-1抑制成纤维细胞α2Ⅰ型胶原的mRNA表达。