Transcriptional Inhibition of Hypertrophic Scars by a Gene Silencer, Pyrrole-Imidazole Polyamide, Targeting the TGF-β1 Promoter
Transcriptional Inhibition of Hypertrophic Scars by a Gene Silencer, Pyrrole-Imidazole Polyamide, Targeting the TGF-β1 Promoter
复制标题
DOI：
10.1038/jid.2011.150
复制
发表时间：
2011-10-01
影响因子：
6.5
通讯作者：
Terui, Tadashi
中科院分区：
文献类型：
--
作者：
Washio, Hisayo;Fukuda, Noboru;Terui, Tadashi
合成的吡咯-咪唑（PI）聚酰胺以高亲和力和特异性结合到双螺旋DNA的小沟上，并抑制相应基因的转录。我们研究了转化生长因子（TGF）-β 1靶向PI聚酰胺（聚酰胺）对大鼠增生性皮肤瘢痕的影响。通过切口在大鼠背侧产生增生性瘢痕。注射FITC标记的聚酰胺以研究其在皮肤中的分布。通过逆转录PCR分析评估TGF-β 1、结缔组织生长因子（CTGF）、1型胶原和纤连蛋白mRNA的表达。纤维化程度和TGF-β 1的表达进行了组织学和免疫组化评价。聚酰胺分布在皮肤细胞的几乎所有的核。切皮后3d TGF-β 1 mRNA表达达到高峰。CTGF和细胞外基质mRNA的表达持续增加，即使在TGF-β 1 mRNA的峰值诱导后。注射聚酰胺完全抑制瘢痕的发展和生长因子和细胞外基质mRNA的诱导。该治疗还显着抑制纤维化变化并减少波形蛋白阳性纺锤形成纤维细胞的数量。注射聚酰胺还减少了大鼠中已形成的增生性瘢痕。因此，TGF-β 1-靶向PI聚酰胺应该是一种可行的基因沉默器，用于增生性瘢痕和瘢痕疙瘩。
Synthetic pyrrole-imidazole (PI) polyamides bind to the minor groove of double-helical DNA with high affinity and specificity, and inhibit the transcription of corresponding genes. We examined the effects of a transforming growth factor (TGF)-beta 1-targeted PI polyamide (Polyamide) on hypertrophic skin scars in rats. Hypertrophic scars were created dorsally in rats by incisions. FITC-labeled Polyamide was injected to investigate its distribution in the skin. Expression of TGF-beta 1, connective tissue growth factor (CTGF), collagen type1, and fibronectin mRNAs was evaluated by reverse transcription PCR analysis. The extent of fibrosis and the expression of TGF-beta 1 were evaluated histologically and immunohistochemically. Polyamide was distributed in almost all nuclei of skin cells. Expression of TGF-beta 1 mRNA reached a peak at 3 days after skin incision. Expression of CTGF and extracellular matrix mRNAs was increased continuously even after the peak induction of TGF-beta 1 mRNA. Injection of Polyamide completely inhibited both the development of scars and the induction of growth factors and extracellular matrix mRNAs. The treatment also markedly inhibited fibrotic changes and reduced the numbers of vimentin-positive spindle-shaped fibroblasts. Injection of Polyamide also reduced established hypertrophic scars in rats. Thus, TGF-beta 1-targeted PI polyamide should be a feasible gene silencer for hypertrophic scars and keloids.