反义纤溶酶原激活物抑制剂1 RNA对家兔动脉粥样硬化形成的影响

The Influence of Antisense Plasminogen Activator Inhibitor-1 RNA on Atherosclerosis in Rabbits

为探讨纤溶酶原激活物抑制剂 1反义RNA对家兔血浆纤溶活性及纤溶酶原激活物抑制剂 1的表达、血脂及对动脉粥样硬化斑块形成的影响 ,通过聚合酶链反应扩增纤溶酶原激活物抑制剂 1第二外显子 ,将聚合酶链反应产物纯化克隆后连入真核细胞表达载体pcDNA3.1,构建纤溶酶原激活物抑制剂 1反义RNA重组质粒。将pcDNA3.1 反义纤溶酶原激活物抑制剂 1重组质粒注射到哈尔滨大白兔腹部皮下组织。通过发色底物法测定家兔血浆组织型纤溶酶原激活物及纤溶酶原激活物抑制剂 1活性变化 ,通过免疫组织化学方法检测组织中纤溶酶原激活物抑制剂 1表达的改变。测定家兔血脂变化 ,病理检测其动脉粥样硬化程度。结果显示 ,应用反义纤溶酶原激活物抑制剂 1RNA重组质粒的家兔血浆纤溶酶原激活物抑制剂 1活性降低 ,组织型纤溶酶原激活物活性升高 ,纤溶酶原激活物抑制剂 1蛋白表达于内皮细胞 ,而在平滑肌细胞中未表达 (动脉粥样硬化对照组中内皮细胞、平滑肌细胞和泡沫细胞内均有表达 ) ;应用反义纤溶酶原激活物抑制剂 1RNA重组质粒的家兔胆固醇和甘油三酯明显低于动脉粥样硬化对照组 (96± 4 2mg L比 12 3± 12mg L ,15± 10mg L比 4 6± 2 9mg L) ,且动脉粥样硬化程度亦轻于后者。以上提示 ,反义纤溶酶原激活物抑制剂

Aim To investigate the role of plasminogen activator inhibitor 1 (PAI 1) antisense RNA in influencing plasma fibrinolytic activity, and regulating the expression of PAI 1 and the process of atherogenesis in rabbits. Methods PAI 1 antisense RNA recombination plasmid was constructed by using eukaryotic cell expression vector pcDNA3.1. Thirty rabbits were classified into 4 groups. Control group (n=7) was fed with a normal diet. Atherosclerosis (As) group (n=7) was fed with a high cholesterol diet. Plasmid treatment group (treatment group, n=9) was fed with a high cholesterol diet and injected with PAI 1 antisense RNA recombination plasmid (20 μg/kg ) into the abdominal subcutaneous tissue weekly. Plasmid group (n=7) was fed with a normal diet and injected with PAI 1 antisense RNA recombination plasmid (20 μg/kg ) into the abdominal subcutaneous tissue weekly. Then the activities of tissue plasminogen activator (t PA), PAI 1, and serumal lipid were detected. PAI 1 protein expression in tissues was tested by immunohistochemistry. Lastly, the extent of atherosclerosis was observed by patho anatomy. Results In plasmid treatment group and plasmid group, the activity of PAI 1 in plasma decreased while that of t PA increased. Although there were no differences between As group and plasmid treatment group in the activities of PAI 1 and t PA at the beginning of the study (P>0.05), in the end the differences were significant (P<0.05). PAI 1 protein only existed in endothelial cells in plasmid treatment group, while in As group, it not only existed in both endothelial cells and smooth muscle cells but also outnumbered the former. Compared with As group, the intima of the arterial vessel was thinner in plasmid treatment group. Plasmid treatment group had a lower lipid count (serum triglyceride and cholesterol) (96±42 mg/L vs 123±12