【摘要】 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-myc AS-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-myc AS-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-myc AS-ODN、PLL载N/P=3:1和N/P=5:1基因复合物的局部转运。24 h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-myc AS-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-myc AS-ODN组cmyc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cmyc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。更多还原

【Abstract】 Objective To investigate the effect of local delivery of phosphorylcholine copolymer MPC30-DEA70carried 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 arteries with ballon catheters.The MPC30-DEA70(N)/FAM-c-myc AS-ODN(P) complexes were transfered into the injured iliac 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 4weeks 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 intima 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 hyperplasia 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.更多还原