纤维蛋白胶联合转染金属蛋白酶组织抑制剂-1预防自体移植静脉再狭窄

Experimental research on the prevention of autologous vein graft restenosis by fibrin glue combined with tissue inhibitor of metalloproteinase-1

目的为减少冠状动脉旁路移植术后移植静脉再狭窄，探讨术中应用纤维蛋白胶联合转染金属蛋白酶组织抑制剂-1（TIMP-1）对自体移植静脉再狭窄的影响及其作用机制。方法将24只新西兰大耳白兔随机分为3组，每组8只，即非支架组（NS组）、纤维蛋白胶外支架组（FS组）、纤维蛋白胶联合转染TIMP—l组（TS／FS组）。建立兔颈外静脉颈总动脉旁路移植术模型。术后28d取出移植静脉桥，进行组织病理分析移植静脉内膜厚度、中膜厚度及内膜面积、中膜面积，用逆转录一聚合酶链反应（RT—PCR）和Westernblot法检测各组移植静脉中TIMP-1基因的表达。结果术后28d，TS／FS组移植静脉内膜厚度及内膜面积分别为（37．98±4．19）μm及（0．557±0．049）mm2，与Ns组的（76．87±4．32）μm、（1．025±0．103）mm2及FS组的（50．28±4．69）μm、（0．743±0．052）mm2比较，明显减少（P〈0．05）；TS／FS组移植静脉中膜厚度及中膜面积分别为（28．45±3．01）μm及（0．458±0．053）mm2，与NS组的（55．98±4．33）μm、（0．944±0．084）mm2及FS组的（36．46±4．36）um、（0．643±0．056）mm。比较，差异有统计学意义（P〈0．05）。与NS组和FS组比较，TS／FS组TIMP-1基因的mRNA和蛋白表达明显增强，差异有统计学意义（P〈0．05）。结论纤维蛋白胶联合血管外膜TIMP—1转染能够实现移植静脉TIMP-1基因的过表达；纤维蛋白胶外支架联合TIMP-1对移植静脉内膜和中膜增生有协同抑制作用。

Objective To investigate the mechanism of attenuation of restenosis by applying fibrin glue combined with tissue inhibitor of metalloproteinase-1 （ TIMP-1 ） locally on vein graft. Methods Twenty-four New Zealand white rabbits were randomly divided into 3 groups with 8 rabbits in each group： non-support group （ NS group）, fibrin glue group （ FS group） and fibrin glue ＋ TIMP-1 group （ TS/FS group）. All animals underwent unilateral jugular vein into common carotid artery interposition grafting. Af- ter operation, vein grafts were harvested at 28th day for histopathologic measurement of intimal thickness, media thickness and intimal areas, media areas, reverse transcription-polymerase chain reaction （RT- PCR） and Western blotting analyses of TIMP-1 gene expression. Results At 28th day after the operation, the mean intimal thickness and intimal areas of the TS/FS group were （37.98 ±4. 19） μm and （0. 557 ±0. 049） mm2 respectively, which were reduced obviously as compared with NS group and FS group （ P 〈 0. 05）. The mean media thickness and media areas in the TS/FS group were （28.45± 3.01 ）μm and （0. 458 ± 0. 053） mm2 respectively, which were decreased statistically as compared with NS group and FS group （ P 〈 0. 05 ）. RT-PCR and Western blotting analysis revealed that the gene expression of TIMP-1 was increased in TS/FS group as compared with NS group and FS group （ P 〈 0. 05 ）. Conclusion The expres- sion of TIMP-1 in vein grafts can be up-regulated by adventitial delivery of TIMP-1. The fibrin glue com- bined with TIMP-1 has synergistic effect on reducing neointimal and media thickness of vein grafts.