二烯丙基三硫化物涂层支架对冠状动脉损伤后内皮结构和功能的影响

Effect of stent coated with diallyl trisulfide on endothelial structure and function after coronary injury: experiment with dogs

目的观察二烯丙基三硫化物涂层支架置入犬冠状动脉后内皮修复、内皮型一氧化氮合酶(eNOS)蛋白质表达和一氧化氮水平的变化,探讨该药物支架对血管内皮结构和功能的影响。方法将8只犬进行冠状动脉左回旋支支架置入术,将二烯丙基三硫化物涂层支架(药物组)和对照支架(对照组)分别置入左回旋支的远端和近端。术后28d处死动物,采用光镜观察内膜增生情况,扫描电镜观察内皮修复情况,用免疫组织化学和Western印迹法测定eNOS蛋白质表达,硝酸还原酶法检测一氧化氮水平。结果术后28d,药物组血管内膜增生(内膜厚度0·12mm±0·05mm,内膜面积1·17mm2±0·25mm2)明显轻于对照组(内膜厚度0·35mm±0·11mm,内膜面积1·81mm2±0·36mm2,P<0·01,P<0·05);血管内膜表面的内皮细胞较对照组分布更紧密,排列更整齐,覆盖更完整;药物组eNOS蛋白质表达量(免疫组化法11·1±1·9,Western印迹法37·5±6·2)明显高于对照组(5·6±0·6,10·4±2·7,均P<0·01);药物组一氧化氮水平(279μmol/L±72μmol/L)明显高于对照组(160μmol/L±57μmol/L,P=0·01)。结论二烯丙基三硫化物涂层支架可加速内皮化、促进eNOS表达和提高一氧化氮水平,表明该药物支架在促进内皮结构修复的同时,实现了内皮功能的恢复和完善。

Objective To investigate the effect of stent coated with diallyl trisulfide on endothelial structure and function after coronary injury. Methods Eight adult dogs were randomly divided into 2 equal groups to undergo implantation of stent coated with diallyl trisulfide （drug group） or stent not coated with diallyl trisulfide （control group） into the distal and proximal ends of the left circumflex coronary arteries respectively. Twenty-eight hours later the dogs were killed. Their coronary arteries with the stents inside were taken out and divided into 4 parts ： 2 parts underwent light microscopy and scanning electron microscopy and 2 parts underwent immunohistochemistry and Western blotting to examine the levels of endothelial nitric oxide synthase （eNOS） and nitrogen monoxide （NO）. Results The thickness of intima of the control group was 0.35 mm ± 0.11 mm, significantly greater than that of the drug group （0.12 mm ± 0.05 mm, P 〈 0. 01 ） ; the area of intima of the control group was 1.81 mm^2 ±0.36 mm^2 , significantly greater than that of the drug group （ 1.17 mm^2 ±0.25 mm^2 , P 〈0.05）. The endothelial cells in the intima of the drug group were tighter and more orderly than those of the control group. Immunohistochemistry showed that the eNOS protein expression of the drug group was 11.1 ± 1.9, significantly higher than that of the control group （5.6 ±0.6, P 〈0.01 ）. Western blotting showed that the eNOS protein expression of the drug group was 37.5 ±6.2, significantly higher than that of the control group （ 10.4 ± 2.7, P 〈 0.01 ）. The NO level of the drug group was 279 μmol/L s 72 μmol/L, significantly higher than that of the control group （60 μmol/L ± 57 72 μmol/L, P = 0.01 ）. Conclusion Stent coated with diallyl trisulfide promotes the endothelialization and expression of eNOS, and raises the NO level. While promoting the recovery of the endothelial structure, stent coated with diallyl trisulfide promotes the recovery of endothelial function.