[诱饵寡脱氧核苷酸对NIH3T3细胞中胶原蛋白基因表达抑制作用的实验研究]
[An experimental study of inhibiting effect of decoy oligodeoxynucleotides on the gene expression of collagen in NIH3T3 cell].
作者信息
Hao Tian-zhi, Liang Huan-ping, Lv Feng-lin, Xu Xiang, Guo Hua, Wang Fu-long, Yang Wen-jun, Luo Yan, Li Lei
机构信息
Department of neurosurgery, Research Institute of surgery, Daping Hospital, Third military Medical University, Chongqing 400042, PR China.
出版信息
Zhonghua Shao Shang Za Zhi. 2003 Jun;19(3):175-8.
OBJECTIVE
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.
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 alpha2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action.
CONCLUSION
Decoy-ODNs could inhibit the mRNA expression of fibroblast alpha2 type I collagen by antagonizing AP-1.
目的
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。
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。
结果
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。
结论
Decoy-ODNs可通过拮抗AP-1抑制成纤维细胞α2Ⅰ型胶原的mRNA表达。
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