转染血小板源生长因子和增殖细胞核抗原反义寡核苷酸抑制移植血管狭窄的实验研究

The study on platelet-derived growth factor and proliferating cell nuclear antigen antisense oligodeoxynucleotides together inhibiting the stenosis of transplanted vascular

目的 探讨联合应用血小板源生长因子(PDGF)和增殖细胞核抗原(PCNA)反义寡核苷酸(AODN)防止移植血管狭窄的作用。方法 将同一只兔的左、右髂外动脉(各1 0cm)对换移植,移植血管用PDGF- AODN和PCNA -AODN转染;移植血管病理切片后测量其内膜厚度、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PCNA阳性细胞数。结果 PDGF- AODN加PCNA- 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PCNA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PCNA -AODN组(P<0. 05 ),PDGF AODN组和PCNA AODN组间差异无显著意义(P>0. 05 )。结论 联合应用PDGF AODN和PCNA- AODN能有效抑制血管平滑肌细胞增殖,阻止内膜增生,防止移植血管狭窄。

Objective To study the effect of platelet-derived growth factor(PDGF)and proliferating cell nuclear antigen (PCNA) antisense oligodeoxynucleotides (AODN) together on inhibiting the proliferation of the stenosis of transplanted vascular. Methods The left and right external iliac arteries(length 1.0 cm) of rabbits were transplanted reciprocally. The transplanted vascular were respectively soaked in lipofiction, PDGF-AODN, PCNA-AODN and PDGF-AODN adding PCNA-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 PCNA′s protein in transplanted vascular wall were counted by S-P immunochemisty.Results The intimal thickness and area, stenosis ratio of transplanted vascular, the number of PDGF and PCNA positive cell in PDGF-AODN adding PCNA-AODN group were significantly lower than those in other group (P<0.01), and that were lower evidently than PDGF-AODN group and PCNA-AODN group. Conclusion PDGF and PCNA antisense oligodeoxynucleotides together could significantly inhibit the proliferation of vascular smooth muscle cell and stenosis of transplanted vascular.