腺病毒介导肿瘤坏死因子样弱凋亡因子转染对大鼠骨髓基质干细胞内皮分化的影响  

Adenovirus-mediated tumor necrosis factor like weak inducer of apoptosis enhanced growth and endothelial differentiation of MSCs in rats

作　　者：于丽[1,2] 孙宏晨[1] 申玉芹[3] 柯小亮[1] 刘霞[4] 苗雷英[4] 

机构地区：[1]吉林大学口腔医院病理科,长春130041 [2]黑龙江省口腔病防治院,哈尔滨150001 [3]吉林大学口腔医院牙周病科,长春130041 [4]吉林大学口腔医院牙体牙髓病科,长春130041

出　　处：《复旦学报（医学版）》2009年第3期261-264,269,共5页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金项目(30672338;30740420551)

摘　　要：目的研究转染腺病毒介导的肿瘤坏死因子样弱凋亡因子(tumor necrosis factor like weak inducer of apoptosis,TWEAK)对大鼠骨髓基质干细胞(bone marrow stromal cells,MSCs)内皮分化功能的影响。方法用密度梯度离心法培养纯化扩增大鼠MSCs细胞,将实验细胞分为转染Ad5CMV-TWEAK的实验组和未经转染的对照组,转染后用生长因子诱导其分化,用CD34和Ⅷ因子相关抗原染色鉴定其分化能力;细胞计数法观测转染TWEAK后MSCs内皮分化细胞增殖的影响;胶面生长实验观察转染TWEAK对MSCs微血管形成能力的影响。结果大鼠MSCs转染TWEAK基因经诱导分化后的CD34和Ⅷ因子相关抗原染色阳性细胞数量增多,与对照组相比有统计学差异(P<0.05)。转染组内皮分化细胞数量增长较快,与对照组相比有统计学意义(P<0.05)。胶面生长实验显示转染TWEAK组细胞微血管新生明显。结论TWEAK基因能促进MSCs向内皮分化,并能促进内皮细胞增殖,在一定的条件下,可促进微血管新生,这为进一步研究其作为诱导体外接种细胞血管发生的基因奠定了基础。Objective To study the effect of the transfection with the recombinant adenovirus containing TWEAK gene on the differentiation of rat bone marrow stromal cells（ MSCs） to endothelial cells. Methods Rat MSCs were isolated by Percoll and cultured by granulocytes adherent. The P3 cells were divided into two groups： experimental and control group. Experiment group were transfected with Ad5CMV-TWEAK. Then inducing culture medium was added to the two groups. Cells differentiation was evidenced by immunohistochemical stain and observed under the microscopy. Cell count showed the effect of TWEAK on the cells. The differention effect of induced cells was analyzed using a fibrin matrix gel assay based on a method that had been previously described. Results There were more CD34 and Factor Ⅶ Ag positive cells in experimental group than control group with the immunohistochemistry stain assay （P〈 0.05）. The results of the cell count showed that there were significance difference between the two groups （P〈0.05）. Many microvasculatures were observed in the experimental group. Conclusions Transfected cells by TWEAK which were induced from MSCs have proliferation and differentiation activity. TWEAK may play a role in the regulation of angiogenesis.

关 键 词：肿瘤坏死因子样弱凋亡因子 骨髓基质干细胞 转染 内皮分化 血管新生 大鼠 

分 类 号：R782.6[医药卫生—口腔医学]