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

1)与对照载体相比，sTNFRI载体转染的HUVEC生长也显著增加。与对照组相比，转染sTNFRI载体的HUVEC在条件培养基中积累的sTNFRI显著增加。转染sTNFRI载体的HUVEC不仅增加了sTNFRI，而且阻止了sTNFRI从TNFRI中脱落。与转染对照载体相比，转染sTNFRI载体的HUVEC也明显阻止了tnf -a诱导的核小体间断裂。2) oxLDL作用于HUVEC后，LOX-1 mRNA水平升高，cpp32样蛋白酶活性升高，诱导细胞凋亡。用硝苯地平预孵养HUVEC，然后用oxLDL孵养，显著抑制LOX-1 mRNA水平和cpp32样蛋白酶活性的增加，以剂量依赖的方式防止(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)本研究报道，在实验性AMI大鼠心肌中直接注射sTNFRI表达质粒DNA可减少梗死面积。用sTNFRI表达质粒DNA处理可降低心肌中tnf - α的生物活性和心肌细胞的凋亡。这些发现提示sTNFRI抗tnf - α治疗可能是治疗AMI的一种新策略。

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.