针对c-fos和c-jun的反义寡核苷酸可抑制大鼠静脉移植模型中的内膜增厚。

Suggs W D, Olson S C, Madnani D, Patel S, Veith F J

Division of Vascular Surgery, Montefiore Medical Center, New York, NY 10467, USA.

Surgery. 1999 Aug;126(2):443-9.

BACKGROUND

C-fos and c-jun are 2 immediate early genes that have been implicated in the stimulation of vascular smooth muscle cell proliferation and migration. In previous experiments in our laboratory with a rat vein graft model a 2- to 3-fold increase of messenger RNA of c-fos and c-jun were noted 1 hour after vein graft perfusion. Because c-fos and c-jun are up-regulated after the perfusion of vein grafts, the purpose of this study was to delineate the temporal expression of c-fos and c-jun protein and to study the effect of antisense oligonucleotides (ASO) to c-fos and c-jun on intimal thickening observed in this model.

METHODS

Sprague-Dawley rats underwent bilateral interposition femoral artery grafts with use of the superficial epigastric vein, which was harvested from 15 minutes up to 2 weeks and analyzed by Western blot for Fos and Jun protein. Additional rats underwent bypasses and at the time of the procedure 1 graft was treated with a pluronic gel containing an ASO to c-fos, c-jun, or sense and the contralateral side was treated with pluronic gel only. The vein grafts were harvested 2 weeks after the procedure and perfusion fixed. After longitudinal sectioning, the intimal and total wall thicknesses were measured in the perianastamotic and midgraft regions by a morphometric digitizing microscope and the statistics were analyzed by a paired Student's t test.

RESULTS

Protein analysis by Western blot showed that c-fos levels rose quickly within 2 hours and leveled at 6 hours 40-fold above basal levels after vein graft perfusion. Similarly, c-jun levels rose 10-fold above basal levels after 15 minutes and peaked at 2 hours 120-fold above basal levels. The treatment of the vein grafts with these ASOs resulted in a reduction of about 30% in the thickness of the intimal layer and the total wall thickness in both the perianastomotic and the midgraft regions, which was statistically significant different from control veins.

CONCLUSION

These results indicate a possible therapeutic role for ASO to immediate early genes in the treatment of vein graft intimal hyperplasia.

背景

C-fos和c-jun是2种即刻早期基因，与血管平滑肌细胞增殖和迁移的刺激有关。在我们实验室先前用大鼠静脉移植模型进行的实验中，静脉移植灌注1小时后，c-fos和c-jun的信使核糖核酸增加了2至3倍。由于静脉移植灌注后c-fos和c-jun上调，本研究的目的是描绘c-fos和c-jun蛋白的时间表达，并研究针对c-fos和c-jun的反义寡核苷酸（ASO）对该模型中观察到的内膜增厚的影响。

方法

Sprague-Dawley大鼠接受双侧股动脉移植，使用腹壁浅静脉，从15分钟至2周采集静脉，通过蛋白质印迹法分析Fos和Jun蛋白。另外的大鼠进行搭桥手术，在手术时，1个移植物用含有针对c-fos、c-jun的ASO或正义链的普朗尼克凝胶处理，对侧用仅含普朗尼克凝胶处理。手术后2周采集静脉移植物并进行灌注固定。纵向切片后，用形态计量数字化显微镜测量吻合口周围和移植物中部区域的内膜和总壁厚度，并通过配对学生t检验进行统计学分析。

结果

蛋白质印迹法分析显示，静脉移植灌注后，c-fos水平在2小时内迅速升高，6小时时达到稳定，比基础水平高40倍。同样，c-jun水平在15分钟后比基础水平升高10倍，在2小时时达到峰值，比基础水平高120倍。用这些ASO处理静脉移植物导致吻合口周围和移植物中部区域的内膜层厚度和总壁厚度减少约30%，与对照静脉相比有统计学显著差异。