PLGA/CS-tPAcDNA静电纺丝血管外支架预防移植静脉再狭窄  

Experimental Study on PLGA/CS-tPAcDNA Electrospun Extravascular Stent for Preventing Restenosis after Transplantation

作　　者：张正刚 伊斯拉木江·吐尔逊 李元明[2] 陈家桦 王磊[2] 刘小斌 Zhang Zhenggang;Yisilamujiang·Tuerxun;Li Yuanming(Department of Cardiac Surgery and Great Vascular Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang 550000,China;Department of Cardiac and Thoracic Vascular Surgery,Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830063,China)

机构地区：[1]贵州医科大学附属医院心血管外科,贵阳550000 [2]新疆医科大学第二附属医院胸心外科,乌鲁木齐830063 [3]华中科技大学同济医学院附属协和医院心血管外科,武汉430022

出　　处：《华中科技大学学报（医学版）》2023年第1期14-17,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：新疆维吾尔自治区自然科学基金资助项目(No.2018D01C226)。

摘　　要：目的通过PLGA/CS-tPAcDNA静电纺丝血管外支架进行t-PA基因局部定位转移,探讨其对移植静脉血栓形成和内膜增生的影响。方法60只兔构建颈外静脉-颈总动脉移植模型,制备PLGA/CS-tPAcDNA纳米粒超细纤维复合膜支架,并随机分为基因治疗组(含t-PA基因PLGA/CS静电纺丝血管外支架,n=30)、对照组(PLGA/CS静电纺丝血管外支架,n=30),分别在术后不同的时间点取样,通过免疫印迹、逆转录聚合酶链反应和底物发色法分别检测移植静脉t-PA活性和基因表达,病理形态学观察移植血管新生内膜增生情况。结果在术后2、5、14、28、60 d,基因治疗组移植静脉可检测到外源性t-PA mRNA的表达,其活性变化分别为(390.42±63.71)、(368.47±54.21)、(275.23±58.32)、(170.75±60.52)和(63.52±23.21)U/g。对照组中对应时间点则t-PA无活性。术后5、14、28、60 d,基因治疗组移植静脉镜下显示新生内膜面积、内膜中膜面积比均明显小于对照组。结论PLGA/CS-tPAcDNA静电纺丝血管外支架可介导t-PA基因局部治疗,可影响移植静脉管腔内血栓形成,抑制移植静脉内膜增殖,继而有效降低移植静脉再狭窄的发生。Objective To investigate the local transfection of t-PA gene by using PLGA/CS-tPAcDNA electrospun external vascular stent,and to explore its effects on thrombosis and intimal hyperplasia of vein grafts.Methods Sixty rabbits were randomly divided into two groups:gene therapy group(PLGA/CS-tPA electrospun vascular stent,n=30)and control group(PLGA/CS electrospun vascular stent,n=30).The expression and activity of t-PA gene were detected by reverse transcriptase polymerase chain reaction(RT-PCR),Western blotting and substrate coloring method,respectively.The neointimal hyperplasia was observed by pathological morphology.Results At day 2,5,14,28 and 60 after the operation,the transcription and protein expression of exogenous t-PAmRNA were detected in gene therapy group.The activity changes of t-PA were(390.42±63.71),(368.47±54.21),(275.23±58.32),(170.75±60.52)and(63.52±23.21)U/g,respectively.No t-PA activity was detected in control group at each time point.At day 5,14,28 and 60 after operation,the neointimal hyperplasia in the gene therapy group were significantly smaller than those in the control group.Conclusion PLGA/CS-tPAcDNA electrospun stent can effectively mediate the local transfection of t-PA gene,inhibit the thrombosis of graft vein,reduce the proliferation of neointima,and effectively prevent the occurrence of restenosis.

关 键 词：组织型纤溶酶原激活因子 基因治疗 静电纺丝血管外支架 静脉移植 再狭窄 

分 类 号：R654.33[医药卫生—外科学]