磷酸胆碱聚合物载反义c—myc寡核苷酸对兔髂动脉球囊损伤后血管狭窄的影响

目的探讨局部转运磷酸胆碱聚合物MPC30-DEA70载反义c—myc寡核苷酸（c—mycAS—ODN）基因复合物对兔髂动脉球囊损伤后血管狭窄的影响。方法40只家兔随机分为对照组、多聚赖氨酸（PLL）组、裸c—mycAS—ODN组、PLL载基因复合物N／P=3：1组和N／P=5：1组，每组各8只。构建兔髂动脉球囊损伤模型，于血管损伤段行PLL、c—mycAS—ODN、PLL载N／P=3：1和N／P=5：1基因复合物的局部转运。24h后应用共聚焦显微镜观察基因复合物在髂动脉的分布情况；4周后苏木精-伊红染色光镜下观察髂动脉的组织形态学变化，包括新生内膜面积（NIA）、中膜面积（MA）和新生内膜／中膜面积比（I／M）；Western blot法检测各组损伤血管处c—myc蛋白的表达情况。结果共聚焦显微镜可见转染N／P=3：1、N／P=5：1基因复合物的血管内膜有均匀的绿色荧光分布。苏木精-伊红染色光镜下对照组、PLL组、裸c—mycAS—ODN组均可见显著的内膜增生，组间比较NIA、I／M无显著性差异（P〉0．05）；与对照组比较，N／P=3：1组和N／P=5：1组NIA、I／M均明显减少（P〈0．05），后两组间比较无显著性差异（P〉0．05）。Western blot显示对照组、PLL组、裸c—mycAS—ODN组c—myc蛋白表达均明显增高，组间比较无显著性差异（P〉0．05）；与对照组比较，N／P=3：1组和N／P=5：1组c—myc蛋白表达明显减少（P〈0．05），后两组间比较无显著性差异（P〉0．05）。结论多聚赖氨酸可促进MPC30-DEA70／c—myc AS—ODN复合物进入髂动脉血管，有效抑制球囊损伤后新生内膜的过度增生，为基因治疗血管内狭窄提供了新型的非病毒类转基因手段。

Effects of phosphorylcholine polymer as a carrier of c-myc antisense oligodeoxynucleotide on the vascular stenosis of iliac arteries in rabbits after balloon injury

Objective To investigate the effect of local delivery of phosphorylcholine copolymer MPC30 - DEA70 carried c - myc AS - ODN on stenosis of iliac artery after balloon injury in rabbits. Methods Fourty rabbits were randomly divided into 5 groups （ n = 8 each） ： control group, PLL group, naked c - myc AS - ODN group, gene complexes groups （N/P =3：1 and N/P = 5：1 ）. The model was established by injury of rabbit iliac a,-with ballon catheters. The MPC30 - DEA70 （ N）/FAM - c - myc AS - ODN （P） complexes were transfered into the injured lilac artery by PLL - coated balloon catheter respectively. The vessel segments were made into fast freezing sections after 24 h and the distribution of gene complexes were observed by confocal laser scanning microscope （ CLSM ）. Rabbits were killed after 4 weeks and morphology of iliac arteries was observed with H -E staining. Neointima area （NIA） , media area （MA） and the ratio of I/M were measured. The expression of c - myc protein in each group was detected by Western blot. Results From CLSM observation, green fluorescence was clearly observed in the vascular infima transfered with gene complexes （N/P =3：1 and N/P =5：1 ）, but the control group, PLL group and naked AS -ODN group were not. All experimental groups showed varying degrees of neointimal byperplasia and vascular stenosis with H - E staining. Compared with the control group, the NIA and I/M in PLL group and naked AS- ODN group had no significant difference （P 〉 0.05 ）. In N/P = 3：1 group and N/P = 5：1 group, the NIA and I/M were reduced significantly （P 〈 0.05）. C - myc protein expressions in control group, PLL group and naked AS - ODN group were obviously increased, but without signifcant difference among different groups （ P 〉 0.05）. Compared with the control group, c - myc protein expressions were significantly decreased in N/P = 3：1 group and N/P = 5 ： 1 group （P 〈 0.05）. Conclusion MPC30 - DEA70 can be used as the carrier of c - myc AS - ODN to rabit iliac artery wtih the aid of PLL, and effectively prevent the stenosis and neointimal hyperplasia atter ballon injury. It provides a new type of nonvirus carrier for gene therapy of vascular stenosis.