Ando Hideyuki, Fukuda Noboru, Kotani Motoko, Yokoyama Shin ichiro, Kunimoto Satoshi, Matsumoto Koichi, Saito Satoshi, Kanmatsuse Katsuo, Mugishima Hideo
Second Department of Internal Medicine, Nihon University School of Medicine, Ooyaguchi-kami 30-1, Itabashi-ku, Tokyo 173-8610, Japan.
Eur J Pharmacol. 2004 Jan 12;483(2-3):207-14. doi: 10.1016/j.ejphar.2003.10.035.
We designed and synthesized a chimeric DNA-RNA hammerhead ribozyme targeting transforming growth factor (TGF)-beta 1 mRNA and found that this ribozyme effectively and specifically inhibited growth of vascular smooth muscle cells. We examined the effects of the chimeric DNA-RNA hammerhead ribozyme targeting TGF-beta 1 mRNA on neointima formation and investigated the underlying mechanism to develop a possible gene therapy for coronary artery restenosis after percutaneous transluminal coronary angioplasty. Expression of mRNAs encoding TGF-beta 1, p27kip1, and connective tissue growth factor (CTGF) in carotid artery increased after balloon injury. Fluorescein-isothiocyanate (FITC)-labeled ribozyme was taken up into the midlayer smooth muscle of the injured carotid artery. Both 2 and 5 mg of ribozyme reduced neointima formation by 65% compared to that of controls. Ribozyme markedly decreased expression of TGF-beta 1 mRNA and protein in injured vessel. Mismatch ribozyme had no effect on expression of TGF-beta 1 mRNA protein in injured vessel. Ribozyme markedly decreased expression of fibronectin, p27kip1, and CTGF mRNAs in injured vessel, whereas a mismatch ribozyme had no effect on these mRNAs. These findings indicate that the chimeric DNA-RNA hammerhead ribozyme targeting TGF-beta 1 mRNA inhibits neointima formation in rat carotid artery after balloon injury with suppression of TGF-beta 1 and inhibition of extracellular matrix and CTGF. In conclusion, the hammerhead ribozyme against TGF-beta 1 may have promise as a therapy for coronary artery restenosis after percutaneous transluminal coronary angioplasty.
我们设计并合成了一种靶向转化生长因子（TGF）-β1 mRNA的嵌合DNA-RNA锤头状核酶，发现该核酶能有效且特异性地抑制血管平滑肌细胞的生长。我们研究了靶向TGF-β1 mRNA的嵌合DNA-RNA锤头状核酶对新生内膜形成的影响，并探究其潜在机制，以期开发一种针对经皮腔内冠状动脉成形术后冠状动脉再狭窄的可能基因治疗方法。球囊损伤后，颈动脉中编码TGF-β1、p27kip1和结缔组织生长因子（CTGF）的mRNA表达增加。异硫氰酸荧光素（FITC）标记的核酶被摄取到损伤颈动脉的中层平滑肌中。与对照组相比，2毫克和5毫克的核酶均使新生内膜形成减少了65%。核酶显著降低了损伤血管中TGF-β1 mRNA和蛋白的表达。错配核酶对损伤血管中TGF-β1 mRNA蛋白的表达没有影响。核酶显著降低了损伤血管中纤连蛋白、p27kip1和CTGF mRNA的表达，而错配核酶对这些mRNA没有影响。这些发现表明，靶向TGF-β1 mRNA的嵌合DNA-RNA锤头状核酶通过抑制TGF-β1以及抑制细胞外基质和CTGF，抑制大鼠颈动脉球囊损伤后的新生内膜形成。总之，针对TGF-β1的锤头状核酶有望成为经皮腔内冠状动脉成形术后冠状动脉再狭窄的一种治疗方法。
