Notch信号和血管内皮生长因子基因对大鼠骨髓间充质干细胞诱导分化后内皮细胞功能的影响  

Effects of Notch signaling pathway and vascular endothelial growth factor gene on the functions of endothelial cells derived from rat bone marrow mesenchymal stem cells

作　　者：晋金兰[1] 庄汉屏[3] 韦建瑞[2] 邓哲彤[1] 张敏[1] 张锐[1] 

机构地区：[1]暨南大学第四附属医院,广州市红十字会医院重症医学科,广东510220 [2]暨南大学第四附属医院,广州市红十字会医院心血管内科,广东510220 [3]中南大学湘雅医院老年医学科

出　　处：《中国危重病急救医学》2012年第3期170-174,共5页Chinese Critical Care Medicine

基　　金：基金项目：中央高校基本科研业务费专项基金资助（21611337）：广东省广州市医药卫生科技项目（201102A213161）;湖南省教育厅科学研究重点项目（湘财[2006]36号）

摘　　要：目的探讨Notch信号和血管内皮生长因子（VEGF165）基因对大鼠骨髓间充质干细胞（MSCs）诱导分化后内皮细胞功能的影响。方法分离、培养大鼠MSCs，用含VEGF165和碱性成纤维细胞生长因子（bFGF）的细胞培养液培养大鼠MSCs2周诱导其向内皮细胞分化；用脂质体将携带有VEGF165基因的质粒转染诱导内皮细胞并对转染效果进行鉴定，逆转录-聚合酶链反应（RT—PCR）检测转染前后细胞上Notch信号受体Notch1和配体Jagged1的表达变化；用γ-内分泌酶抑制剂L-685458阻断细胞Notch信号通路的转导，划痕实验检测细胞迁移能力；将细胞接种在半固体培养基上，观察其形成毛细血管样结构的能力。结果转染VEGF165基因的内皮细胞上表达有VEGF165mRNA，说明实验成功地将VEGF165基因导入诱导后内皮细胞中。转染VEGF，65基因后，细胞上Notch信号配体JaggedlmRNA表达增强（1．08±0．01比1．01±0．02。P〈0．01），NotchlmRNA表达无明显变化（0．60±0．02比0．59±0．01，P〉0．05）；细胞的迁移能力增强（划痕空白处细胞个数：46．45±4．46比41．61±1．42，P〈0．05），形成毛细血管样结构能力无明显变化（细胞分级：3．00±0．89比2．00±0．89，P〉0．05）。内皮细胞转染VEGF165基因后，以γ-内分泌酶抑制剂L-685458阻断细胞Notch信号通路的转导，则细胞迁移能力（划痕空白处细胞个数：51．72±3．47比46．45±4．46）和形成毛细血管样结构能力（细胞分级：4．17±0．75比3．00±0．89）均进-步增强（均P〈0．05）。结论转染VEGF165基因可增强大鼠MSCs诱导分化内皮细胞的功能，在此基础上阻断Notch信号通路转导可进-步增强细胞功能。Objective To explore the effects of Notch signaling pathway and the vascular endothelial growth factor （VEGF165） gene on the functions of endothelial cells derived from rat bone marrow mesenehymal stem cells （MSCs）. Methods Isolated and cultivated rat bone marrow MSCs in vitro, then the cells were treated by VEGF165 and basic fibroblast growth factor （bFGF） for 2 weeks to induce them to differentiate into endothelial cells. The gene of VEGF165 was transfeeted into differentiated endothelial cells to promote the functions of the ceUs. The receptor Notehl and the ligand Jaggedl of the Notch signaling were detected by reverse transeripfion-polymerase chain reaction （RT-PCR） before and after the transfeetion. 3,-seeretase inhibitor L-685458 was used to block Notch pathway. Migration ability of cells was detected by scarification test. Cells were inoculated on semisolid gel to study their ability of forming capillary-like structure. Results After transfeetion, VEGF165mRNA could be detected on the differentiated endothelial cells. The expression of Jaggedl mRNA was up regulated （ 1.08 ± 0.01 vs. 1.01 ±0.02, P〈0.01 ） and there was no change in Notehl mRNA（0.60 ± 0.02 vs. 0.59 ± 0.01, P〉0.05 ）. The ability of migration was enhanced （number of cells on the scratched area：46.45 ±4.46 vs. 41.61± 1.42,P〈0.05）, and the ability of forming capillary-like structure on semisolid gel showed no change （cells classification： 3.00 ± 0.89 vs. 2.00 ± 0.89,P〉0.05）. After the tranfection, using the γ-secretase inhibitor L-685458 to block the Notch signaling transduetion, the ablility of migration of the differentiated endothelial cells （ number of cells on the scratched area： 51.72± 3.47 vs. 46.45 ± 4.46, P〈0.05 ）, and that of forming capillary-like structure （cells classification： 4.17 ±0.75 vs. 3.00 ±0.89, P〈0.05 ）, was also further enhanced. Conclusion Transfeetion of the gene of VEGF165 into the differentiated endothelial cells can reinforce the function of thes

关 键 词：NOTCH信号 血管内皮生长因子 骨髓间充质干细胞 内皮细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]