P21基因转染治疗移植静脉内膜增生的潜在临床价值研究

目的:研究复制缺陷性腺病毒携带p21基因体外孵育转染治疗移植静脉内膜增生(IH)的可行性及潜在临床价值。方法:采用直接连接法将p21基因装入5型腺病毒,得Adp21。实验动物分3组(n=36):p21基因转染组(T)、药物治疗组(D)、对照组(C),T组移植静脉段取材后置于高滴度的病毒工作液(2×10~8pfu/ml)中,0℃～4℃下体外孵育转染45min,转染后将该静脉段经端端吻合间置于同侧颈总动脉;D组动物于术前1周给抗IH药西拉普利,C组动物无特殊处理,后2组移植静脉段取材后置于普通培养液45min(0℃～4℃下),同法吻合移植。于术后不同时间段取材,印迹杂交及免疫组化法鉴定p21基因的表达,HE及V.G染色观察移植静脉段的IH情况。结果:T组术后3d即有p21表达,另两组不表达。D组的I/M在术后3d至术后3w小于C组,有显著差异(P<0.05),至4w时D组的I/M已接近C组,无显著差异。3d时D组I/M小于T组,但1w后各时间段的情况则相反,T组I/M小于D组,均有显著差异(P<0.05,具体数值见表-1)。结论:体外孵育法可有效实现Adp21转染移植静脉段,静脉移植入体内后能有效表达转染的目的基因,功能蛋白p21能减轻移植静脉IH,其效果优于抗IH药西拉普利,有重要的潜在临床意义。

Study on the Potential Clinical Significance of Intimal Hyperplasia Inhibitionafter Autologous Vein - to - Artery Transplantation by Overexpression of Ade-noviral Mediated p21 Gene Transfer

Objective: To investigate the possibility of intimal hyperplasia (IH) inhibition after in - vitro incubated autolo-gous vein - to - artery transplantation by overexpression of replication - deficient adenovirus mediated p21 gene transfer and its perspective clinical significance. Methods: The recombinant replication - deficient adenovirus Adeno - p21 harboring the desired gene was constructed by direct ligation between p21 gene and type 5 adenovirus. The animals were randomized into three groups (n =36): Adp21 transfection group (T group), cilazapril treated group (D group) and the normal control group (C group). In the T group, a segment of the right external jugular vein was harvested and placed in the high liter (2