输注表达VEGF的成纤维细胞和rAAV-VEGF基因对大鼠脑缺血保护作用的比较研究  

An investigation on the efficacy of transferring fibroblasts expressing VEGF and delivering rAAV-VEGF into rat cerebral ischemia model

作　　者：邵林华[1] 孙青芳[2] 卞留贯[2] 陈亦华[1] 程振宇[1] 郑龙[1] 俞建华[1] 张晨冉[2] 顾吕伟[2] 

机构地区：[1]浙江省金华市人民医院神经外科,浙江金华321000 [2]上海瑞金医院神经外科,上海200025

出　　处：《中风与神经疾病杂志》2011年第8期712-714,共3页Journal of Apoplexy and Nervous Diseases

基　　金：浙江省卫生厅科研计划(2007B221);金华市科技局科研计划基金资助课题(2007-03-044)

摘　　要：目的比较定向移植输注转染外源性VEGF(血管内皮细胞生长因子)基因的成纤维细胞(VEGF-成纤维细胞)和原位注射rAAV(重组腺相关病毒)载体介导的VEGF基因对大鼠缺血性脑保护的疗效,从而寻找出适合临床的安全、有效的基因治疗方法。方法用线栓加环扎法建立SD大鼠大脑中动脉持续性闭塞(MCAO)模型。30只雄性SD大鼠随机分为假手术组、移植输注组、直接注射组、MCAO组和正常对照组。其中,移植输注组和直接注射组于术后24h内将VEGF-成纤维细胞和rAAV1-VEGF基因从大鼠脑缺血边缘区注入。结果移植VEGF-成纤维细胞和原位注射rAAV-VEGF基因均能减少脑梗死体积,且移植VEGF-成纤维细胞的作用明显优于原位注射基因,VEGF-成纤维细胞移植14d后大鼠脑内表达VEGF的阳性细胞数明显多于原位注射rAAV-VEGF治疗组。结论移植VEGF-成纤维细胞比原位注射rAAV-VEGF治疗基因对脑缺血更具保护作用。Objective To compare the different effect of transferring fibroblasts expressing vascular endothelial growth factor（VEGF） and delivering recombinant adeno-associated virus mediated vascular endothelial growth factor（rAAV-VEGF） gene into brain of rats on ischemic brain tissues.Methods We established a rat model of permanent middle cerebral artery occlusion（MCAO） by nylon suture embolization and cerclage.The fibroblasts expressing VEGF or rAAV-bFGF gene was directly injected through skull into the ischemic lesion within 24h after MCAO.On 14d after the fibroblasts or gene was transferred,the rats were killed separately.VEGF protein in brain tissues was detected by immunohistochemistry stain,and infarct size was analyzed by TTC staining.Results From observing the VEGF protein level in the ischemic penumbra and the size of infarct,it was found that an obvious increase of VEGF protein level and infarct volume was observed in the therapeutic group when compared to the MCAO and sham-operated group（P〈0.05）,and it was more significant in the therapeutic group by transferring fibroblasts expressing VEGF than by rAAV-VEGF group（P〈0.05）.Conclusion Our results indicate that the therapy by transferring fibroblasts expressing VEGF to the ischemic cerebrovascular diseases is a safe and effective gene transferring approach that may be suitable for clinical status.

关 键 词：脑缺血 成纤维细胞 血管内皮细胞生长因子 血管生成 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]