明胶涂层涤纶片携带组织型纤溶酶原激活因子基因定位转移可能影响兔左房血纤溶的活性吗?  

Does gelatin coating dacron patch containing source of tissue-type plasminogen activator gene transfer have effects on plasminogen activity in rabbit left atrium?

作　　者：刘小斌[1] 张凯伦[2] 蒋雄刚[2] 夏家红[2] 向道康[1] 李岑[1] 

机构地区：[1]贵州省人民医院心血管外科,贵州省贵阳市550001 [2]华中科技大学同济医学院附属协和医院心血管外科,湖北省武汉市430020

出　　处：《中国组织工程研究与临床康复》2010年第12期2141-2144,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30571838);课题名称:tPAcDNA基因瓣膜瓣膜预防机械瓣膜置换术后血栓形成的实验研究;贵州省优秀科技教育人才省长专项基金(黔省专合字200763号);课题名称:RNAi干扰沉默组织因子基因预防移植静脉再狭窄的实验研究~~

摘　　要：背景:机械瓣膜置换后的抗凝治疗一直是困扰心血管外科医生的难题,因此研制一种不需终生抗凝的机械瓣膜具有重要的现实意义。目的:观察明胶涂层涤纶片(Dacron片)携带组织型纤溶酶源激活因子(tPA)基因左心房局部定位转移对兔左房血纤溶活性的影响,为tPA基因瓣膜的研制奠定基础。方法:48只新西兰大耳白兔随机分为3组:转基因组左心耳植入携带tPAcDNA的明胶涂层的Dacron涤纶片;载体对照组左心耳植入携带载体DNA的明胶涂层的Dacron片,空白对照组左心耳植入不携带任何基因的明胶涂层的Dacron片。术后3,14d取心肌组织,RT-PCR检测左房局部心肌组织tPAmRNA转录水平,Western-Blot和免疫组化方法检测外源性tPA蛋白表达;术后3,14d取兔左房血和外周静脉血,底物发色法检测其tPA活性。结果与结论:术后3,14d,RT-PCR可检测到局部心肌tPAmRNA表达,Western-Blot可检测到tPA蛋白表达,免疫组化见阳性心肌细胞胞浆呈黄褐色,术后3,14d,转基因组左房血tPA活性明显高于各对照组(P<0.05)。3组术后外周血tPA活性差异无显著性意义。说明使用明胶蛋白涂层方法能够将tPA基因成功的转移到左心房,使之在局部持续表达、分泌有活性的tPA蛋白,从而使左房血纤溶活性增强。BACKGROUND：Anticoagulant therapy after cardiac valve replacement puzzles cardiovascular surgeons;therefore,it has great value to develop mechanical valves that do not need anticoagulant therapy.OBJECTIVE：To evaluate the effects of locally applied gelatin coating dacron patch containing source of tissue-type plasminogen activator（t-PA）gene transfer on plasmingen activity in left atrium and to provide a basis for construction of tPAcDNA gene valve.METHODS：A total of 48 New Zealand white rabbits were divided into 3 groups,including transfected gene group（implant gelatin coating dacron patch containing tPAcDNA in left atrium）,empty vector group（implant gelatin coating dacron patch containing empty vector）and blank group（implant gelatin coating dacron patch without gene）.The specimens were harvested at 3 and 14 days after operation.The expression of tPA gene of some myocardium tissue in left atrium muscle was detected by RT-PCR and Western-Blot essay.The tPA activity of plasmingen tissues in left atrium and peripheral blood was measured by chromogenic substrate essay.RESULTS AND CONCLUSION：At 3 and 14 days after operation,RT-PCR and Western-Blot confirmed the expression of tPAmRNA and the presence of tPA protein at the site of gene transfer,which was stained yellow brown.The tPA activity in experimental group was significantly higher than that in control groups（P 0.05）.The differences of tPA acitive have no significance.Gelatin coating dacron patch is feasible to be the carrier of gene transfection.Locally applied tPA gene can increase tPA activity in left aterium.

关 键 词：组织型纤溶酶原激活因子 基因 明胶涂层 左心房 纤溶活性 生物材料 

分 类 号：R318[医药卫生—生物医学工程]