转染PDGF和c-myc反义寡核苷酸抑制移植血管狭窄研究

The study on platelet-derived growth factor and c-myc together inhibiting the stenosis of transplanted vascular

目的:探讨血管移植后联合应用血小板源生长因子(Platelet-derived growth factor,PDGF)和原癌基因c-myc反义寡核苷酸(Antisense oligodeoxynucleotides,AODN)抑制血管平滑肌(Vascular smooth muscle cell,VSMC)增殖和防止移植血管狭窄的作用。方法:选48只新西兰雄性白兔,随机分为对照组、PDGF-AODN组、c-myc-AODN组和PDGF-AODN+c-myc-AODN组,每组12只;将同一只兔的左、右髂外动脉(各1.0 cm)对换移植,移植血管用PDGF-AODN和c-myc-AODN液浸泡,血管吻合口用AODN液浸泡过的缝线吻合;取移植血管制片显微镜下观察内膜增生情况,计算机图像分析测量其内膜厚度、内膜面积和移植血管狭窄率,并分别用原位杂交组织化学和免疫组织化学染色计数PDGF和c-myc阳性细胞数。结果:PDGF-AODN+c-myc-AODN组的移植血管内膜厚度、内膜面积、管腔狭窄程度和PDGF阳性细胞及c-myc阳性细胞数均显著低于对照组(P<0.01),而且亦明显低于单独转染PDGF-AODN组或c-myc-AODN组(P<0.05),PDGF-AODN组和c-myc-AODN组间差异无显著意义(P>0.05)。结论:联合应用PDGF-AODN和c-myc-AODN能有效抑制VSMC增殖,阻止内膜增生,防止移植血管狭窄,为利用反义核酸技术联合调控多个基因防止移植器官内血管狭窄奠定了基础。

Objective：To study the effect of platelet-derived growth factor（PDGF）and c-myc antisense oligodeoxynucleotides（AODN） together on inhibiting the proliferation of vascular smooth muscle cells（ VSMC） and the stenosis of transplanted vascular.Method：To select 48 male Zelanian rabbits and were randomly divided into four groups（n=12,in each group）：control group,PDGF-AODN group,c-myc-AODN group,PDGF-AODN adding c-myc-AODN group.The left and right external iliac arteries（length 1.0cm） of rabbits were transplanted reciprocally.The transplanted vascular were respectively soaked in lipofiction,PDGF-AODN,c-myc-AODN and PDGF-AODN addingj c-myc-AODN solution about 20 minute,the vascular anastomotic were sutured by 8/0 suture of soaked in AODN solution.Four weeks later,the specimens were harvested for microscopy.The pathological morphology of transplanted vascular were observed under microscope（HE）.The intimal thickness and area,stenosis ratio（%） of transplanted vascular were calculate and analysed statistically among group by computer system.The number of positive cells of PDGF＇s mRNA in transplanted vascular wall were counted with in situ hybtidzatian histo-cytochemistry and the number of positive cells of c-myc＇s protein in transplanted vascular wall were counted by S-P immunochemisty.Result：The intimal thickness and area,stenosis ratio of transplanted vascular,the number of PDGF and c-myc positive cell in PDGF-AODN adding c-myc-AODN group were significantly lower than those in other group（P0.01）,and that were lower evidently than PDGF-AODN group and c-myc-AODN group.Conclusion：PDGF and c-myc antisense oligodeoxynucleotides together could significantly inhibit the proliferation of VSMC and stenosis of transplanted vascular. The present study lay the foundation for fuetur making use of oligodeoxynucleotides technology to prevent and treat vascular stenosis in transplanted organ.