目的 探讨可溶性支架转染c-myc反义寡核苷酸的可行性及其对静脉桥内膜增生的作用,以利在分子水平对静脉桥再狭窄进行防治.方法 新西兰大耳白兔随机分成5组,每组10只.建立兔颈外静脉颈总动脉旁路移植术模型.合成c-myc寡核苷酸,处理可溶性支架,依组别不同,在静脉移植时管腔内分别置人:(1)空白对照;(2)单纯可溶性支架;(3)反义寡核苷酸可溶性支架;(4)正义寡核苷酸可溶性支架;(5)不匹配寡核苷酸可溶性支架;静脉移植后7、28和90d取出静脉桥.行HE及弹力纤维染色,计算内膜、中膜厚度及内膜/中膜比值;利用免疫组化方法,检测各组静脉移植物内c-myc基因及PCNA的表达;采用原位杂交及激光灰度扫描,检测各组静脉桥中c-myc基因mRNA表达水平.结果 静脉移植后7、28和90d反义寡核苷酸可溶性支架组静脉桥内膜增生程度显著降低;c-myc及PCNA蛋白的表达显著低于同时间点其他各组;c-myc基因mRNA的表达显著低于同时间点其他各组.结论 可溶性支架可实现c-myc基因反义寡核苷酸的转染,可溶性支架转染c-myc基因反义寡核苷酸可显著抑制静脉桥c-myc及PCNA基因的表达,预防静脉桥再狭窄.
Objective A rabbit model of common carotid artery grafted by external jugular veins was used. Vein grafts were transferred by c-myc anfisense oligedecxynucleotides(ODN) and carried by soluble stent. Try to find a new approach to prevent veins graft failure on molecular basis, netlmds New Zealand rabbit were randomly divided into five groups, 10 animals each. Pretreating the soluble stents with synthesized c-myc ODN, following stents: (1) control group; (2) soluble stent; (3) soluble stent with senseODN; (4) soluble stent with anfisense-ODN; (5) soluble stent with mismatch-ODN were put into the vein graft during end to end anastomosis. After 7, 28 and 90 days of operation, vein grafts were harvested. HE and ET stain were made aim to calculate the extent of intima hypeiplasia. The expression of c-myc and PCNA were identified by immtmchemistry methods. Situ hybridization and Northern Bloting were made to assess the expression of c-myc mRNA quantitatively and semi-quantitatively in veins. Results In the vein grafts of 7,28,90 days post-operation of soluble stent antisense-ODN group ①Intima hypeiplasia were inhibited significantly compare with other four groups. ②The expression of c-myc and PCNA were inhibited significantly compare with other four groups. ③C-myc mRNA expression level was significantly Lower than the other four groups. Conclusion Soluble stent can transfer c-myc ODN effectively. C-myc antisense-ODN transferred by soluble stent can inhibit the expression of c-myc and PCNA, can inhibit intima hyperplasia of vein graft significantly, thus preventing the stenosis of vein graft.