不同途径导入rAAV-VEGF_(165)基因对大鼠缺血性脑保护的比较研究  

Experimental studies on the different routes of transfer rAAV-VEGF_(165) Gene into Ischemic Rat Brain

作　　者：饶宜光 褚晓凡 付学军 郑佩娥[2] 马可夫 黄莲婵 邹良玉 

机构地区：[1]暨南大学第二临床医学院深圳市人民医院神经内科,深圳518020 [2]暨南大学医学院病理教研室 [3]暨南大学第二临床医学院深圳市人民医院特诊科,深圳518020

出　　处：《中国神经精神疾病杂志》2007年第4期213-219,共7页Chinese Journal of Nervous and Mental Diseases

摘　　要：目的比较不同途径(直接注射、脑脊液及静脉内)导入重组腺相关病毒介导的血管内皮细胞生长因子基因(recombinant adeno-associated virus mediated vascular endothelial growth factor165 gene,rAAV-VEGF165)治疗大鼠局灶性脑缺血的疗效,寻找出适合临床、安全、有效的基因导入方法。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大鼠随机分为直接注射组、脑脊液导入组、静脉导入组、MCAO组和假手术组,其中治疗组于术后24h内将rAAV-VEGF165基因从大鼠脑缺血边缘区、小脑延髓池或股静脉内注入。术后14d取脑,免疫组织化学染色检测脑缺血边缘区VEGF的表达和微血管的密度、HE染色观察脑组织坏死情况、4%TTC染色检测脑梗死体积。结果与单纯梗死组和假手术组相比较,治疗组脑缺血边缘区VEGF表达水平明显升高、微血管密度显著增高(P<0.01),脑组织坏死情况明显减轻,脑梗死体积明显缩小(P<0.05);其中以前2个指标在直接注射和脑脊液导入组比静脉导入组更为显著(P<0.05)。结论直接注射和脑脊液导入比静脉内导入rAAV-VEGF165基因转染到大鼠缺血脑组织的效果更优,而且通过脑脊液导入可能是较为合适有效的途径。Objective To compare the different effect of transferring recombinant adeno-associated virus mediated vascular endothelial growth factor165 gene （ rAAV-VEGF165 ） into brain of rats on ischemic brain tissues by three pathways ： via cerebrospinal fluid （CSF） ,via intravenous route and directly injecting through skull into the ischemic lesion. From observing the expression level of vascular endotheial growth factor （VEGF） , the microvascular density in the ischemic penumbra （IP） and the size of infarct, we explore a safe and effective gene transferring approach that suitable for clinical status and provide experimental data for the VEGF gene therapy of the ischemic cerebrovascular diseases. Methods We established a rat model of permanent middle cerebral artery occlusion （MCAO） by nylon suture embolization and cerclage, the rAAV-VEGF165 gene was injected through the cerebellomedullais cisterna , the thigh vein and directly injected through skull into the ischemic focal within 24 hours after MCAO. On the day 14th after the rAAV-VEGF165 gene was transfered , the rats were killed separately. Brain tissues was stained by HE, infarct size was analysed by TTC staining, and the VEGF protein and the microvascular density were detected by immunohistochemistry stain in IP. Results High levels of VEGF protein expression, a significant increase in the number of vessels （P 〈 0..01 ） and a significant reduction in the ratio of infarct volume and necrosis zone （P 〈 0. 05 ） in the gene therapeutic groups were observed when compared to the MACO without transfection group and sham-operated group. In three gene therapeutic groups, we also found the effect was better in the directly injected and CSF injected groups than that in the intravenous groups. Conclusions Our results indicate that the effect wasbetter in the directly injected and CSF injected groups than that in the intravenous groups. And the CSF route is a safe and effective gene transfering pathway of approach that suitable for clinical statu

关 键 词：脑缺血 重组腺相关病毒介导血管内皮细胞生长因子基因 血管生成 基因治疗 

分 类 号：R743.3[医药卫生—神经病学与精神病学]