反义N-ras1基因对血管损伤后平滑肌细胞增殖的影响

Influence of antisense N ras 1 gene on smooth musele cell proliferation after arterial injury

为了防止血管损伤后狭窄或球囊成形术后再狭窄，观察反义Ｎ－ｒａｓ１基因对血管损伤后血管平滑肌细胞（ＶＳＭＣ）增殖的影响。方法实验动物随机分为基因治疗组、空载体对照及正义Ｎｒａｓ１重组质粒对照组。将实验血管进行血管造影、病理形态学检测、提取血管组织ｍＲＮＡ与Ｎ－ｒａｓ１探针进行Ｎｏｒｔｈｅｒｎ印迹杂交，应用Ｗｅｓｔｅｒｎ技术检测ｒａｓ癌基因族表达产物ｐ２１蛋白。结果血管造影显示，基因治疗组（治疗后４周）血管直径０．８０±０．１０（ｍｍ）；空载体对照组０．３５±０．１３（ｍｍ），两组差异有非常显著意义（Ｐ＜０．０１）；组织学检查显示治疗组新生内膜面积（治疗后４周）０．４３±０．０５ｍｍ２，空载体对照组０．８２±０．０３ｍｍ２，两组差异有非常显著意义（Ｐ＜０．０１）。治疗组血管ｒａｓｍＲＮＡ表达明显减少，且表达蛋白ｐ２１也明显受到抑制。

Objectives To develop anew way to prevent the process of restenosis after arterial injury,we transfered the recombinant antisense N ras 1 gene to the iliac arteries of rabbits after injury and studied the influence of gene therapy with antisense N ras1on the vascular segment.Methods The constructed recombinant antisense N ras 1 gene to deliver into the iliac arteries of rabbits after injury.The rabbits were randomly divided into gene therapy group,vector group,and sense N ras 1 control.The segments of treatment rabbits were analysed by angiography of vessels,pathological study,Northern blotting,and Western blotting technique.Results The minimum angiographic lumen diameter measured during gene therapy and at follow up,for the gene therapy group(at 4 weeks)was 0.80±0.10mm and the vector contro l0.35±0.13mm( P <0.01).Histologically,the area of neointima measured,for gene the therapy group(at4weeks)was0.43±0.05mm 2,and the vector control group 0 82±0.03mm 2( P < 0.01). By using Northern blotting and Western blotting technique,we found that cell's ras mRNA of the vessel segments of gene local delivery was significantly lower than that of the control.Cell's p21 was significantly lower than that of the control.Conclusions The antisense N ras 1 inhibites VSMC proliferation.The result implicates the potential value in future gene therapy of restenosis.