反义c-myc和增殖细胞核抗原抑制移植血管狭窄研究

c-myc and proliferating cell nuclear antigen antisense oligodeoxynucleotides together inhibiting the stenosis of transplanted vessels

目的探讨血管移植后联合应用c-myc和增殖细胞核抗原（PCNA）反义寡核苷酸（AODN）抑制血管平滑肌增殖和防止移植血管狭窄的作用。方法选40只新西兰雄性白兔，随机分为对照组（A组）、c-myc-AODN组（B组）、PCNA-AODN组（C组）和c-myc-AODN加PCNA-AODN组（D组），每组10只；将同一只兔的左、右髂外动脉（各1．0cm）对换移植，移植血管用c-myc-AODN和PCNA-AODN液浸泡转染；术后4周时B超观察移植血管通畅情况和内径，同时取移植血管制片显微镜下测量其内膜厚度、内膜面积和移植血管狭窄率，并用免疫组织化学染色计数c-myc和Pc-NA阳性细胞数。结果D组的移植血管内膜厚度、内膜面积、管腔狭窄程度和c-myc阳性细胞及PCNA阳性细胞数均显著低于A组（P〈0．01），而且亦明显低于B组或C组（P〈0．05）；其内径比A组（P〈0．01）、B组和C组显著增大（P〈0．05）；B组和C组间差异无统计学意义（P〉0．05）。结论联合应用c-myc-AODN和PCNA-AODN能有效抑制VSMC增殖，阻止内膜增生，防止移植血管狭窄。

Objective To study the inhibitory effect of c-myc and proliferating cell nuclear antigen （PCNA） antisense oligodeoxynucleotides （AODN） together on the proliferation of vascular smooth muscle cells （VSMC） and the stenosis of transplanted vessels. Methods Forty male Zelanian rabbits were randomly divided into 4 groups （ n = 10 in each group） ： control group （ group A）, c-myc-AODN group （group B）, PCNA-AODN group （group C）, c-myc-AODN ＋ PCNA-AODN group （group D）. The left and right external iliac arteries （length 1.0 cm） of rabbits were transplanted reciprocally. The transplanted vessels were respectively soaked with c -myc. AODN and PCNA -AODN and transfected about 20 min. Four weeks later, the diameter of transplanted vessels was measured by B ultrasound apparatus and specimens were harvested for microscopic examination. The pathological morphology of transplanted vessels was observed under a microscope （HE staining）. The intimal thickness and area, stenosis ratio （%） of the transplanted vessles were calculated and analyzed statistically among the groups by computer system. The number of c-myc and PCNA positive cells in transplanted vascular wall was counted by S-P immunochemisty. Results The intimal thickness and area, stenosis ratio of the transplanted vessels, the number of c-myc and PCNA positive cells in group D were significantly deceased as compared with groups A, B and C （P 〈0.01 or P 〈0.05）. The diameter of the transplanted vessels in group D was increased significantly as compared with that in groups A, B and C （P 〈 0.05）. Conclusion c-myc and PCNA antisense oligodeoxynucleotides together could significantly inhibit the proliferation of VSMC and stenosis of transplanted vessles.