超声靶向转纳米tPA基因预防兔动脉粥样硬化血栓形成

Targeting transfection of nano t-PA gene by ultrasound to prevent thrombosis induced by rabbit artery atherosclerosis

目的观察构建的纳米白蛋白超声微泡载t-PA基因靶向转染骨骼肌和局部血管预防兔髂动脉粥样硬化血栓形成的效果。方法制备兔髂动脉粥样硬化血栓形成模型;构建高表达白蛋白纳米t-PA基因超声微泡靶向载体并在超声波介导下靶向转染骨骼肌和局部血管壁,动态观察对局部血栓形成的作用。免疫组化法检测转染骨骼肌组织t-PA抗原表达;ELISA法检测血t-PA和D-二聚体含量变化,观察t-PA活性。结果超声介导靶向转染基因获得局部骨骼肌和血管组织有效表达tPA抗原并伴有血tPA和D-二聚体含量增高、tPA活性增强,有效预防了兔实验性髂动脉粥样硬化血栓形成。结论成功建立了白蛋白纳米tPA基因超声微泡载体系统,为血栓相关性疾病预防提供了实验依据。

[Objective] To observe the effect of constructed ultrasound microbubbles crosslinked to albumin nanoparticles packaged with t-PA gene plasmid on thrombosis induced by rabbit artery atherosclerosis. [ Methods ] A rabbit model of iliac artery atherosclerosis and thrombosis was prepared. T-PA gene plasmid was constructed and packaged with bovine serum albumin nanoparticles. This t-PA gene nanoparticles was crosslinked to an ultrasound microbubbles prepared with sucrose and bovine serum albumin to form a targeting vector for t-PA gene transfection and the skeleton muscle tissue around the local arteries was targeted]y transfected with the introduction of ultrasonic wave （1 kHz, 1.5 W/cm2, l0 min）. The expression of t-PA in local muscle was detected with indirect immunohistochemical method and blood t-PA level and D-dimer content were detected with enzyme linked immunosorbent assay before, 1 and 2 weeks after operation. [ Results ] The effective expression of t-PA could be seen in local muscle tissue and on artery wall with increases in blood t-PA and D-dimer content. The thrombosis induced by artery injury and local atherosclerosis was prevented 2 weeks after operation. [ Conclusion] The albumin nanoparticle-microbubble-t-PA gene targeting vector was constructed successfully. This provides a experimental basis for the prevention of thrombosis-related diseases.