缺氧诱导人脑动静脉畸形血管内皮细胞生长因子表达  

Hypoxia-induced expression of VEGF gene in endothelial cells cultured from patients with cerebral arteriovenous malformation

作　　者：赵明光[1] 陈游力[1] 高永中[2] 浦佩玉[2] 宋振全[1] 魏学忠[1] 

机构地区：[1]中国人民解放军沈阳军区总医院神经外科,辽宁沈阳110016 [2]天津医科大学总医院神经外科,天津300052

出　　处：《中国微侵袭神经外科杂志》2006年第6期261-263,共3页Chinese Journal of Minimally Invasive Neurosurgery

摘　　要：目的探讨缺氧条件下体外培养的人脑动静脉畸形(cAVM)血管内皮细胞中血管内皮细胞生长因子(VEGF)基因表达与细胞超微结构变化。方法采用组织块贴壁法对cAVM血管内皮细胞进行培养和形态学观察。采用免疫组化方法检测细胞中第八因子相关抗原(FⅧ-RA)的表达,采用RT-PCR技术观察静态和缺氧状态下2 h、4 h、8 h血管内皮细胞VEGF mRNA表达,采用酶联免疫吸附实验(ELISA)检测每组细胞上清液中VEGF蛋白含量;在透射电镜下观察细胞超微结构的变化。结果体外培养的活细胞具有单层“卵石样”排列的典型特征,95%以上的细胞FⅧ-RA染色阳性。血管内皮细胞VEGF蛋白和mRNA表达在缺氧2 h开始升高(P<0.05),并持续至缺氧8 h(P<0.01);细胞线粒体丰富,核糖体及粗面内质网逐渐增多。结论缺氧可能在基因转录水平诱导VEGF表达,后者通过自分泌途径刺激cAVM血管内皮细胞增殖。Objective To investigate the gene expression of vascular endothelial growth factor （VEGF） and ultraslructural changes in in vitro cultured endothelial cells from cerebral arteriovenous malformation （cAVM） patients under hypoxic condition. Methods The endothelial cells in the blood vessel with cAVM were grown by explant culture techniques and observed morphologically after culturing for 60 hours and purifying using a cell-scraping technique. The expression of factor Ⅷ-relative antigen （FⅧ-RA） in the cultured cells was observed by immunocytochemistry. The levels of VEGF mRNA in the cells and VEGF protein in the cell supernatant were determined by RT-PCR analysis and ELISA respectively when they were exposed to hypoxic conditions for 2h, 4h and 8h, or maintained in basal condition. Ultrastructural changes of the cells were also observed by electron microscopic technique. Results Under inverted microscope the cultured cells showed contact inhibition and a cobblestone-like appearance. More than 95% of the cells were stained strongly with antibodies against F Ⅷ-RA. Levels ofVEGF mRNA and protein started to increase at 2h （P〈0.05）, and the increase persisted for 8h （P〈0.01） after these cells were exposed to hypoxia, and the mitochondrion, ribosome, rough endoplasmic reticulum and extracellular matrix became obviously abundant. Conclusion The gene expression of VEGF in endothelial cells of the vessel with cAVM in response to hypoxia may provide a novel mechanism that the proliferation of endothelial cells can be regulated in an autocrine manner, and this may play an important role in the pathogenesis and development of vascular malformations in the central nervous system.

关 键 词：颅内动静脉畸形 内皮细胞 缺氧 血管内皮生长因子A 

分 类 号：R743.4[医药卫生—神经病学与精神病学] R364.4[医药卫生—临床医学]