Development of gene therapy for the vascular proliferative diseases by ribozyme targetting TGF-β mRNA

我们设计了针对TGF-β_1 mRNA的锤头核酶，用于切割合成的底物RNA。我们研制的DNA-RNA嵌合核酶能显著抑制自发性高血压大鼠血管平滑肌细胞（VSMC）的生长。DNA-RNA嵌合核酶抑制SHR VSMC TGF-β_1 mRNA和蛋白表达。DNA-RNA嵌合核酶显著抑制shrr - sp系膜细胞中TGF-β_1、纤维连接蛋白、胶原I型mrna的表达。在体内实验中，fitc标记的核酶在主动脉和肾脏中有效分布。DNA-RNA嵌合核酶在体内显著抑制SHR-SP患者肾动脉肥大和肾小球硬化。我们将DNA-RNA嵌合核酶应用于猪血管成形术后冠状动脉狭窄，通过确定疗效、剂量和治疗周期，开发DNA-RNA嵌合核酶TGF-β_1作为动脉增殖性疾病和肾血管性硬化的基因治疗方法。

We designed hammerhead ribozyme targetting TGF-β_1 mRNA which cleaved the synthetic substrate RNA. We developed DNA-RNA chimeric ribozyme, which significantly inhibited growth of vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR). DNA-RNA chimeric ribozyme inhibited TGF-β_1 mRNA and protein in VSMC from SHR. DNA-RNA chimeric ribozyme significantly suppressed expression of TGF-β_1, fibronectin, collagen type I mRNAs in mesangial cells from SHR-SP. In vivo experiments, FITC-labeled ribozyme was efficiently distributed in aorta and kidney. DNA-RNA chimeric ribozyme considerably inhibited hypertrophy of renal capirally artery and gromerulosclerosis in SHR-SP in vivo. We are appling DNA-RNA chimeric ribozyme on coronary stenosis after angioplasty in pig, by which efficiency, dosage, and period of treatment will be determined to develop the DNA-RNA chimeric ribozyme to TGF-β_1 as a gene therapy for arterial proliferative diseases and gromerulosclerosis.