【摘要】 目的探讨局部应用5-氟尿嘧啶(5-fluorouracil,5-FU)对动脉缺损静脉移植后内膜增生的影响。方法64只5月龄雄性新西兰大白兔,体重2.8～3.0kg,随机分为A、B、C、D4组(n=16)。游离左侧颈总动脉,中间切除长约1cm,建立动脉缺损模型。切取右侧颈外静脉3cm,将切取的静脉远近端倒置后用9-0无损伤缝线端端吻合于动脉缺损。吻合结束后A、B、C组动物分别用经50.0、25.0、12.5mg/mL5-FU浸泡的脑棉(大小为12mm×30mm×1mm)完全包裹移植静脉外膜,共5次,每次1min,D组用生理盐水同法处理。分别于术后1、2、4、6周切取移植静脉,行HE、Masson染色观察移植静脉管壁组织学变化,增殖性细胞核抗原(proliferating cell nuclear antigen,PCNA)免疫组织化学染色、TUNEL标记染色观察移植静脉平滑肌细胞增殖和凋亡情况,透射电镜观察细胞超微结构变化。结果术后1、2、4、6周,HE染色、Masson染色、PCNA免疫组织化学染色均示A、B组移植静脉内膜增厚明显小于C、D组,术后1、2、4周A、B组增殖细胞少于C、D组。内膜厚度、内膜增生程度、管腔狭窄程度:术后1周A组分别为(12.69±1.68)μm、0.73±0.05、0.025±0.003;B组分别为(17.52±2.01)μm、0.86±0.06、0.027±0.004;C组分别为(21.92±1.85)μm、1.06±0.09、0.036±0.006;D组分别为(26.45±3.86)μm、1.18±0.08、0.041±0.005。术后2周A组分别为(24.61±2.91)μm、0.86±0.06、0.047±0.003;B组分别为(37.28±2.78)μm、1.17±0.09、0.060±0.004;C组分别为(46.52±2.25)μm,1.44±0.08、0.073±0.003;D组分别为(52.07±3.29)μm、1.45±0.05、0.081±0.006。术后4周A组分别为(61.09±6.84)μm、1.38±0.08、0.106±0.007;B组分别为(63.61±8.25)μm、1.40±0.07、0.107±0.010;C组分别为(80.04±7.65)μm、1.64±0.07、0.129±0.011;D组分别为(84.45±9.39)μm、1.68±0.10、0.139±0.014。术后6周A组分别为(65.27±5.25)μm、1.46±0.07、0.113±0.005;B组分别为(65.82±7.12)μm、1.45±0.05、0.112±0.011;C组分别为(84.45±9.39)μm、1.69±0.09、0.135±0.007;D组分别为(87.27±9.96)μm、1.76±0.05、0.140±0.012。上述指标及术后1、2、4周细胞增殖指数均为A、B组小于C、D组,且差异有统计学意义(P<0.05);术后1、2周,内膜、中膜细胞凋亡指数A、B组均大于C、D组,差异有统计学意义(P<0.05)。透射电镜示A、B组相对于C、D组粗面内质网、高尔基体、核糖体等合成细胞器含量明显减少。结论局部应用5-FU可有效抑制移植静脉内膜增生。更多还原

【Abstract】 Objective To assess the effect of topical application of 5-fluorouracil(5-FU) on intimal hyperplasia in rabbit vein graft.Methods Sixty-four male New Zealand white rabbits, aged 5 months and weighing 2.8-3.0 kg, were randomly divided into group A, B, C, and D(n=16 rabbits per group).Artery defect model was established by cutting about 1 cm artery from the middle part of the dissociated left common carotid artery.A section about 3 cm was cut from the right external jugular vein, and the harvested vein was inverted and end-to-end anastomosed to the artery defect with 9-0 non-traumatic suture.After anastomosis, the extima of the grafted veins in group A, B, and C was completely wrapped with cotton sheet(12 mm × 30 mm × 1 mm in size) immersed by 5-FU at a concentration of 50.0, 25.0, and 12.5 mg/mL, respectively, and each vein was treated 5 times(1 minute at a time).In group D, the extima of the graft veins was treated with normal saline instead of 5-FU.The grafted veins were obtained 1, 2, 4, and 6 weeks after operation, HE staining and Masson staining were preformed for histological changes of grafted vein wall, proliferating cell nuclear antigen(PCNA) immunohistochemistry staining and TUNEL labeling staining were conducted for proliferation and apoptosis of smooth muscle cell of the grafted vein, and transmission electron microscope observation was performed for cellular ultrastructure.Results The HE staining, Masson staining, and PCNA immunohistochemistry staining showed that the thickness of intima in group A and B was obviously less than that in group C and D at 1, 2, 4, and 6 weeks after operation, and the proliferation cells in group A and B were less than that in group C and D at 1, 2, and 4 weeks after operation.The thickness of the intima, the degree of intima hyperplasia, the degree of vessel lumen stenosis of four groups at different time points were as follows:at 1 week after operation, group A(12.69 ± 1.68) μm, 0.73 ± 0.05, 0.025 ± 0.003, group B(17.52 ± 2.01) μm, 0.86 ± 0.06, 0.027 ± 0.004, group C(21.92 ± 1.85) μm, 1.06 ± 0.09, 0.036 ± 0.006 and group D(26.45 ± 3.86) μm, 1.18 ± 0.08, 0.041 ± 0.005;at 2 weeks after operation, group A(24.61 ± 2.91) μm, 0.86 ± 0.06, 0.047 ± 0.003, group B(37.28 ± 2.78) μm, 1.17 ± 0.09, 0.060 ± 0.004, group C(46.52 ± 2.25) μm, 1.44 ± 0.08, 0.073 ± 0.003, and group D(52.07 ± 3.29) μm, 1.45 ± 0.05, 0.081 ± 0.006;at 4 weeks after operation, group A(61.09 ± 6.84) μm, 1.38 ± 0.08, 0.106 ± 0.007, group B(63.61 ± 8.25) μm, 1.40 ± 0.07, 0.107 ± 0.010, group C(80.04 ± 7.65) μm, 1.64 ± 0.07, 0.129 ± 0.011, and group D(84.45 ± 9.39) μm, 1.68 ± 0.10, 0.139 ± 0.014;at 6 weeks after operation, group A(65.27 ± 5.25) μm, 1.46 ± 0.07, 0.113 ± 0.005, group B(65.82 ± 7.12) μm, 1.45 ± 0.05, 0.112 ± 0.011, group C(84.45 ± 9.39) μm, 1.69 ± 0.09, 0.135 ± 0.007, and group D(87.27 ± 8.96) μm, 1.76 ± 0.05, 0.140 ± 0.012.Group A and B were inferior to group C and D in terms of the above three parameters and cell proliferation index 1, 2 and 4 weeks after operation(P < 0.05).Group A and B were superior to group C and D in terms of cell apoptosis index of intima and media 1 and 2 weeks after operation(P < 0.05).Transmission electron microscope observation showed that the synthetic cell organelles such as rough endoplasmic reticulum, golgi apparatus, and ribosome in group A and B were obviously less than those in group C and D(P < 0.05).Conclusion Topical application of 5-FU can effectively inhibit intima hyperplasia of the vein grafts.更多还原