【摘要】 目的:构建高表达腺病毒感染的体外三维血管生成模型,为精确研究某个基因在血管新生中的功能提供技术平台。方法:制备空载腺病毒和cdc42高表达腺病毒。用空载腺病毒感染人微血管内皮细胞(hum an m icrovascu lar endothelial cell,HMVEC),然后用感染过的HMVEC构建体外血管生成三维模型,同时用未感染的HMVEC构建对照组模型,观察腺病毒感染本身对血管新生模型的生长有无影响。用cdc42高表达腺病毒感染HMVEC并构建体外血管新生模型,对照模型采用空载腺病毒感染的细胞构建,验证由腺病毒介导的基因高表达在模型发展过程中维持的时间。结果:空载腺病毒感染模型的平均出芽长度(average length of sprouts,ALS)在24、48、72 h时与非感染模型相比有显著性差异(P<0.01),在96、120 h时与非感染模型无显著性差异(P>0.01)。cdc42高表达腺病毒感染的模型在模型构建后1周内保持了高表达。结论:成功地使用腺病毒作为载体诱导了某个基因在体外三维血管生成模型中的高表达,为精确研究某个基因在血管新生中的功能提供技术平台。更多还原

【Abstract】 Objective To construct an in vitro three-dimensional angiogenic model infected by over expression of adenovirus,thus to set up a platform for studying some specific gene in angiogenesis.Methods Blank adenovirus and cdc42 over expression adenovirus were prepared.Blank adenovirus was used to infect primary human microvascular endothelial cell(HMVEC),then the in vitro three-dimensional angiogenic model was constructed with the infected HMVECs,as well as the control model with noninfected HMVECs.The effect of infection on the growth of the angiogenic model was observed.cdc42 over expression adenovirus was used to infect primary HMVEC to construct the in vitro three-dimensional angiogenic model with the infected HMVECs,simultaneously blank adenovirus was used to infect the control model with HMVECs.The time the over expression lasted in the model developed was measured by reverse transcription PCR.Results There was a significant difference between average length of sprouts(ALS) of blank adenovirus infected models and ALS of noninfected models at the time points 24,48,72h(P<0.01) repectively.There was no significant difference between ALS of the two models at the time points of 96 and 120h(P>0.01).The over expression of cdc42 was maintained in the model at least for a week.Conclusions Adenovirus has been successfully used as a vector to induce over expression of some specific gene in the in vitro three-dimensional angiogenic model.A platform for studying accurate role of some specific gene in angiogenesis has been established.更多还原