pcDNA3.1(-)/tPA-交联明胶微球复合物体外基因转染的实验研究  

The Gene Transfection Study of pcDNA3.1(-)/tPA-Cross Linking Gelatin Microspheres Complexes in Vitro

作　　者：祖育昆[1] 张凯伦[1] 蒋雄刚[1] 郭超[1] 厉泉[1] 

机构地区：[1]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：《微循环学杂志》2008年第2期1-3,7,F0003,共5页Chinese Journal of Microcirculation

基　　金：国家自然科学基金(No.30571838)

摘　　要：目的:检测交联明胶微球结合和缓释pcDNA3.1(-)/tPA的能力及其转染体外培养人脐静脉内皮细胞的瞬时转染效率。方法:采用紫外分光光度法检测交联明胶微球结合pcDNA3.1(-)/tPA的最大包封率和体外缓释规律;采用免疫荧光染色法检测pcDNA3.1(-)/tPA-交联明胶微球复合物缓释的pcDNA3.1(-)/tPA转染内皮细胞的瞬时转染效率。结果:交联明胶微球能有效结合pcDNA3.1(-)/tPA,最大包封率为62.75±3.31%;且在体外能持续缓释质粒DNA超过4周。pcDNA3.1(-)/tPA-交联明胶微球复合物缓释的pcDNA3.1(-)/tPA能被转移进入人脐静脉内皮细胞中并有效表达,瞬时转染效率为12.74±2.53%。结论:交联明胶微球能良好结合和缓释pcDNA3.1(-)/tPA,并能成功进行体外基因转染。Objective:To investigate the ability of incorporating and delay releasing pcDNA3.1(-)/tPA from the cross linking gelatin microspheres prepared in previous research,and evaluate the possibility and transient efficiency of pcDNA3.1(-)/tPA,released from the cross linking microspheres,being transferred into human umbilical vein endothelial cells in vitro.Method:Ultraviolet spectrophotometry was adopted to investigate the ability of incorporating pcDNA3.1(-)/tPA from the cross linking gelatin microspheres by measuring the maximum entrapment radio.It was also adapted to investigate the time profile of delay releasing pcDNA3.1(-)/tPA.Immunofluorescence staining was used to evaluate the possibility and transient efficiency of gene transferred in vitro.Results:The maximum entrapment radio of incorporating pcDNA3.1(-)/tPA was 62.75±3.31%.pcDNA3.1(-)/tPA could be persistently released from the cross linking gelatin microspheres for over 4 weeks in vitro.pcDNA3.1(-)/tPA released from the cross linking gelatin microspheres could be effectively transferred into HUVECs with efficiency of 12.74±2.53%.Conclusion:These studies demonstrated the cross linking gelatin microspheres had good ability of incorporating and delay releasing pcDNA3.1(-)/tPA,and plasmid DNA could be successfully transferred into HUVECs in vitro.

关 键 词：体外基因转染 组织型纤溶酶原激活物 实验研究 明胶微球 PLASMINOGEN 复合物 ACTIVATOR 交联 

分 类 号：R767.91[医药卫生—耳鼻咽喉科] R743.33[医药卫生—临床医学]