【摘要】 目的　探索转基因疗法和激光疗法防治移植静脉远期再狭窄的可行性及作用机制。方法　建立兔颈外静脉颈总动脉移植模型 ,分为 (1)对照组 ,(2 )绿色荧光蛋白 (GFP)基因转染组 ,(3)p5 3基因转染组 ,(4)低能量激光照射组 ,(5 )p5 3基因转染并低能量激光照射组。术后 4周 ,免疫组织化学方法检测外源p5 3基因的表达及增殖细胞核抗原 (PCNA) ,应用DNA片段末端标记法 (TUNEL)标记凋亡细胞。HE、Masson及维多利亚兰染色后 ,应用计算机图像分析系统检测移植静脉内膜、中膜增生情况。结果　术后 4周 ,与对照组相比 ,GFP基因转染组移植静脉血管平滑肌细胞 (VSMC)增殖率、凋亡率差异无显著性 ,移植静脉内膜和中膜厚度无明显变化 ;p5 3基因转染组VSMC增殖率降低 6 1% ,凋亡率增加 2 5 % ,移植静脉内膜和中膜厚度分别减少 6 0 %、33% ,内膜厚度 中膜厚度比值 (I M)减少 37% ;应用低能量激光照射组VSMC增殖率降低 41 5 % ,细胞凋亡率增加 40 9% ,移植静脉内膜和中膜厚度分别减少 5 8 5 %、18 0 % ,I M比值减少 47 2 % ;转染p5 3基因同时应用低能量激光照射组VSMC增殖率较对照组降低6 1 7% ,细胞凋亡率增高 47 0 % ,移植静脉内膜和中膜厚度分别减少 6 9 7%、44 4% ,I M比值减少44 5 %。结论　转染野生型p5 3基因和低?更多还原

【Abstract】 Objective: To evaluate the possibility of using trans-gene therapy and laser therapy in management of late vein graft failure, as a new strategy for further experimental study and clinical application. Methods: In the rabbit a model of jugular veins grafted into the carotid arteries was established. Animals were randomly divided: control group, p53 gene transfected group, GFP gene transfected group, low energy laser irradiated group, p53 gene transfected and low energy laser irradiated group. Four weeks after the operation, p53 protein and proliferating cell nuclear antigen (PCNA) were localized by immunohistochemical procedure, apoptosis was evaluated through terminal d-UTP nick end-labeling (TUNEL). The area of neointima and medial were calculated from sections stained with HE, Masson and Victoria blue by computer image analysis. Results: After the adenovirus delivery of the wild type p53 gene, p53 protein was found to be over-expressed in the VSMC of the vein graft. Four weeks after the operation, compared with the control group, the proliferation rate of the VSMC decreased 61%, while the apoptosis rate increased 25%. The thickness of the intima and media decreased 60% and 33%, respectively. The ratio of intima thickness/media thickness (I/M) decreased 37%. However, 4 weeks after the adenovirus delivery with over-expression of GFP gene in the vein graft by the same way, compared with the control group, there was no significant difference in the proliferation and apoptosis rate of the VSMC, the thickness of the intima and media differed little either. In the low energy laser irradiated group, 4 weeks after the operation, compared with the control group, the proliferation rate of the VSMC decreased 42.5%, the apoptosis rate increased 40.9%, the thickness of the intima and media decreased 58.5% and 18.0%, respectively. The I/M ratio decreased 47.2%. In the vein graft treated with wild-type p53 gene transfection and low energy laser irradiation, 4 weeks after the operation, compared with the control group, the proliferation rate of the VSMC decreased 61.7%, the apoptosis rate increased 47.0%, the thickness of the intima and media decreased 60.7% and 44.4%, respectively. The I/M ratio decreased 44.5%. Conclusions: Wild-type p53 gene transfection and low energy laser irradiation of the vein [FQ(4。29(+219mm,0mm)-W] 作者单位:100853 北京,解放军总医院心外科(迟立群、李功宋、高长青、朱朗标),激光科(刘凡光、顾瑛) *现在首都医科大学附属北京安贞医院心外科博士后流动站 100029graft can inhibit the proliferation of the VSMC, promote the apoptosis of the VSMC, and prevent the hyperplasia of both neointima and media. The results highlight the potential therapeutic benefit of wild-type p53 gene transfection and/or low energy laser irradiation in treating late vein graft failure.更多还原