Pelisek Jaroslav, Fuchs Alexandra T, Kuehnl Andreas, Tian Wen, Kuhlmann Michael T, Rolland Pierre H, Mekkaoui Choukri, Gaedtke Lars, Nikol Sigrid
Department of Cardiology and Angiology, Westfaelische Wilhelm University, Muenster, Germany.
J Gene Med. 2006 Jul;8(7):835-44. doi: 10.1002/jgm.905.
Restenosis is still a significant clinical problem limiting the long-term therapeutic success following balloon dilation or stent implantation. New approaches are necessary inhibiting neointima formation and simultaneously promoting re-endothelialization. Therefore, long-term therapeutic effects of adventitial liposome-mediated C-type natriuretic protein (CNP) gene and CNP peptide applications in a porcine model for restenosis post-angioplasty were investigated.
For in vitro applications, primary cultures of porcine vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) were used. Gene transfer was performed with cationic lipid DOCSPER [1,3-dioleoyloxy-2-(N5-carbamoylspermine)propane]. In vivo treatment of pig femoral arteries was adventitial using a needle injection catheter following balloon angioplasty. Arteries were investigated by angiography, Evan's blue staining, histomorphometry, immunohistochemistry, PCR and RT-PCR.
Using CNP gene transfer in vitro, 29.4+/-7.2% reduction of cell proliferation in VSMCs was observed. In ECs, the CNP gene did not compromise cellular growth. For the CNP peptide the optimal concentration was 1 mM with 50.7+/-11.3% reduction of VSMC proliferation and 12.1+/-5.3% enhancement of growth of ECs. Three weeks following application in vivo complete re-endothelialization was observed in all treated groups. At 3 months significant reduction of neointima formation was observed using CNP gene vs. CNP peptide (85.9+/-7.8% vs. 63.3+/-27.6% reduction, P<0.05) compared to control treatment.
Periadventitial liposome-mediated CNP gene transfer in vivo resulted in a significant long-term reduction of neointima formation without compromising endothelial repair and was superior over single CNP peptide administration. Advantages of CNP are its physiological origin and simultaneous inhibition of VSMC proliferation and promotion of EC growth.
再狭窄仍是一个严重的临床问题，限制了球囊扩张或支架植入术后的长期治疗效果。需要新的方法来抑制内膜增生，同时促进内皮再形成。因此，研究了外膜脂质体介导的C型利钠肽（CNP）基因和CNP肽在猪血管成形术后再狭窄模型中的长期治疗效果。
体外应用时，使用猪血管平滑肌细胞（VSMC）和内皮细胞（EC）的原代培养物。采用阳离子脂质DOCSPER[1,3-二油酰氧基-2-（N5-氨甲酰基精胺）丙烷]进行基因转移。球囊血管成形术后，使用针式注射导管对外膜猪股动脉进行体内治疗。通过血管造影、伊文思蓝染色、组织形态计量学、免疫组织化学、PCR和RT-PCR对动脉进行研究。
体外使用CNP基因转移时，观察到VSMC中细胞增殖减少29.4±7.2%。在EC中，CNP基因不影响细胞生长。对于CNP肽，最佳浓度为1 mM，VSMC增殖减少50.7±11.3%，EC生长增强12.1±5.3%。体内应用三周后，所有治疗组均观察到完全内皮再形成。与对照治疗相比，在3个月时，使用CNP基因与CNP肽相比，内膜增生显著减少（减少85.9±7.8%对63.3±27.6%，P<0.05）。
体内外膜脂质体介导的CNP基因转移可显著长期减少内膜增生，而不影响内皮修复，且优于单次给予CNP肽。CNP的优点在于其生理来源以及同时抑制VSMC增殖和促进EC生长。
