Development of new gene therapy for treatment of atherosclerosis using anti-apoptotic genes

1)转染sTNFRI载体的人脐静脉内皮细胞的生长速度也明显高于对照组。与对照组相比，转染sTNFRI载体的HUVEC条件培养液中sTNFRI的积累量显著增加。转染sTNFRI载体的人脐静脉内皮细胞不仅增加了sTNFRI，而且阻止了sTNFRI从TNFRI中脱落。转染sTNFRI载体的人脐静脉内皮细胞与对照载体相比，其诱导的核小体断裂也被显著阻止。2)与oxLDL共同孵育人脐静脉内皮细胞后，LOX-1基因表达水平和CPP32蛋白水解酶活性显著升高，并诱导细胞凋亡。在oxLDL孵育前用硝苯地平预先孵育HUVEC可显著抑制LOX-1mRNA水平和CPP32样蛋白酶活性的升高，并呈剂量依赖关系。这些结果表明，硝苯地平通过降低LOX-1mRNA水平和CPP32样蛋白酶活性来阻断导致内皮细胞凋亡的自杀途径。3)我们报道了将sTNFRI表达载体DNA直接注射到心肌中减少实验性大鼠急性心肌梗死的梗塞范围。经sTNFRI表达载体DNA处理后，心肌组织中的肿瘤坏死因子-α生物活性降低，心肌细胞凋亡率降低。提示sTNFRI抗肿瘤坏死因子-α治疗可作为治疗急性心肌梗死的新策略。

1) Growth of HUVEC transfected with sTNFRI vector also increased significantly compared to those transfected with control vector. Accumulation of sTNFRI significantly increased in conditioned medium from HUVEC transfected with sTNFRI vector compared to those transfected with control vector. HUVEC transfected with sTNFRI vector not only increased sTNFRI but also prevented shedding of sTNFRI from TNFRI. The TNF-a-induced internucleosomic fragmentation was also significantly prevented in HUVEC transfected with sTNFRI vector compared to those transfected with control vector.2) The incubation of HUVEC with oxLDL increased LOX-1 mRNA levels and CPP32-like protease activity, and induced apoptosis. Preincubation of HUVEC with nifedipine before incubation with oxLDL significantly suppressed the increase in LOX-1 mRNA levels and CPP32-like protease activity, preventing (7)___ in a dose-dependent manner. These results suggest that nifedipine blocks the suicide pathway leading to the apoptosis of endothelial cells by decreasing LOX-1 mRNA levels and CPP32-like protease activity.3) Here we report that direct injection of a sTNFRI expression plasmid DNA to the myocardium reduces infarct size in experimental rat AMI. Treatment with sTNFRI expression plasmid DNA reduced the TNF-alpha bioactivity in the myocardium and the apoptosis of cardiomyocytes. These findings suggest that the anti-TNF-alpha therapy by sTNFRI can be a new strategy for treatment of AMI.