高糖环境对骨形态发生蛋白-2、胰岛素样生长因子-1转染大鼠骨髓间充质干细胞基因表达的影响  

Effect of transfected bone morphogenetic protein-2 gene and insulin-like growth factor gene on gene expression of Wistar rat mesenchymal stem cells

作　　者：王亮[1] 邢德国[2] 吴建军[1] 刘中浩[2] 田克立[3] 宫明智[2] 

机构地区：[1]山东大学,济南250033 [2]山东大学第二医院骨外一科 [3]山东大学分子生物学实验室

出　　处：《中华实验外科杂志》2011年第4期615-617,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：山东省科技攻关计划资助项目（2007GG20002034）

摘　　要：【摘要】目的观察骨形态发生蛋白-2（BMP-2）、胰岛素样生长因子-1（IGF-1）在高糖环境下能否转染大鼠骨髓间充质干细胞（BMSC），转染成功后目的基因表达。方法腺病毒载体介导大鼠BMP-2和IGF-1基因高糖环境下转染Wistar大鼠BMSC，应用逆转录一聚合酶链反应（RT-PCR）和酶联免疫吸附试验（ELISA）定量检测基因复制及表达。结果高糖环境下BMP-2、IGF-1基因各自及共同转染BMSC后，BMSC成活，BMP-2基因表达的产物峰值出现在转染后72h，BMP-2组灰度值为（1．86±0．06），产物浓度为（58．60±1．61）μg/L，BMP-2＋IGF-1组灰度值为（2．374-0．34），产物浓度为（67．61±1．91）μg/L，两组差异均有统计学意义（P〈0．05）；IGF-1基因表达的产物峰值出现在转染后48h，IGF．1组灰度值为（1．48±O．03），产物浓度为（65．17±3．64）μg／L，BMP．2＋IGF-1组灰度值为（1．93±0．17），产物浓度为（73．94±4．30）μg／L，两组差异均有统计学意义（P〈0．05）。结论高糖环境下BMP-2、IGF-1可转染BMSC，且转染后基因表达产物量显著增加。BMP-2、IGF-1基因在基因表达上有协同作用。Objective To investigate whether bone morphogenetic protein-2 （BMP-2） and insu- lin-like growth factor-1 （ IGF-1 ） in high glucose can be transfected into Wistar rat mesenchymal stem cells, and the gene expression after transfection. Methods Adenovirus-mediated BMP-2 and IGF-1 genes were transfected into the Wistar rat mesenchymal stem cells. Quantitative reverse transcription-polymerase chain reaction （RT-PCR） and enzyme linked immunosorbent assay （ELISA） were used to detect the gene repli- cation and expression. Results After the transfeetion of adenovirus-mediated BMP-2 and（or） IGF-1 gene into the cells in high glucose, all of bone marrow stromal cells survived, bone marrow stromal ceils could express the target gene product with the peak of BMP-2 gene product expression at in 72nd h after transfec- tion. The grey degree and gene product concentration of BMP-2 in BMP-2 group were （ 1.86 ± 0. 06） and （58.60 ± 1.61 ） p,g/L, and those in BMP-2 ＋ IGF-1 group （2. 37 ± 0. 34） and （67.61 ± 1.91 ） μg/L, respectively （ all P 〈 0. 05 ）. The peak of IGF-1 gene expression occurred at 48th h after transfection. The grey degree and gene product concentration of IGF-1 in IGF-1 group were （ 1.48 ± 0. 03 ） and （65.17 ± 3.64） μg/L, and those in BMP-2 ＋ IGF-1 group were （1.93 ±0. 17） and （73.94 ±4. 30） p,g/L, respec- tively （ all P 〈 0. 05 ）. Conclusion Adenovirus-mediated BMP-2 and IGF-1 gene could be transfected the Wistar rat bone marrow stromal cells in high glucose. The gene products after transfection were increased significantly. There was a synergistic effect on gene expression between BMP-2 and IGF-1.

关 键 词：骨形态发生蛋白-2 胰岛素样生长因子-1 高糖 骨髓间充质干细胞 

分 类 号：R587.2[医药卫生—内分泌]