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

1)与转染对照载体的HUVEC相比，转染sTNFRI载体的HUVEC的生长也显着增加。与用对照载体转染的那些相比，用sTNFRI载体转染的HUVEC的条件培养基中sTNFRI的积累显着增加。用sTNFRI载体转染的HUVEC不仅增加了sTNFRI而且还阻止了sTNFRI从TNFRI中脱落。与转染对照载体的HUVEC相比，转染sTNFRI载体的HUVEC也显着阻止了TNF-α诱导的核小体间断裂。2) HUVEC与oxLDL的孵育增加了LOX-1 mRNA水平和CPP32样蛋白酶活性，并诱导细胞凋亡。在与 oxLDL 一起孵育之前，将 HUVEC 与硝苯地平预孵育可显着抑制 LOX-1 mRNA 水平和 CPP32 样蛋白酶活性的增加，从而以剂量依赖性方式防止 (7)___。这些结果表明，硝苯地平通过降低 LOX-1 mRNA 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) 在此，我们报告直接向心肌注射 sTNFRI 表达质粒 DNA 可以减少实验性大鼠 AMI 中的梗死面积。 sTNFRI表达质粒DNA处理降低了心肌中TNF-α的生物活性和心肌细胞的凋亡。这些发现表明，sTNFRI 的抗 TNF-α 疗法可能成为治疗 AMI 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.