高糖对骨形态发生蛋白-2、胰岛素样生长因子-1基因转染大鼠骨髓基质干细胞增殖的影响  

Effects of transfection of bone morphogenetic protein-2 and insulin-like growth factor-1 gene on the proliferation of bone marrow mesenchymal stem cells with high glucose condition

作　　者：吴建军[1] 邢德国[2] 王亮[1] 田克立[3] 刘中浩[2] 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3期465-467,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：目的 观察高糖环境下骨形态发生蛋白-2（BMP-2）和胰岛素样生长因子-1（IGF-1）基因转染大鼠骨髓基质干细胞（BMSC）后BMSC的增殖.方法 用Ad-BMP-2和Ad-IGF-1转染大鼠BMSC,Wester blot检测蛋白表达.噻唑蓝（MTT）比色法及流式细胞术检测细胞增殖.结果 Western blot检测到转染组细胞中有目的 蛋白表达.MTT结果显示第5天细胞增殖能力达到高峰,5 d光吸收值：A～E组分别为0.324±0.027、0.319±0.017、0.622±0.028、0.626±0.020、0.778±0.031.流式细胞术结果显示A～E组细胞处于增殖期的比重分别为23.92±3.07、23.51±2.11、34.37±6.85、35.04±1.45、42.56±1.15.结论 高糖环境下BMP-2和IGF-1基因转染均能促进BMSC增殖,联合转染对BMSC增殖有协同效应.Objective To observe the proliferation of bone marrow stromal cell （BMSC） transfected by bone morphogenetic protein-2 （BMP-2） and insulin-like growth factor-1 （ IGF-1 ） gene under high glucose condition.Methods Ad-BMP-2 and Ad-IGF-1 transfected rat BMSC,protein expression of BMSC were detected by Western blotting analysis.Methyl thiazol tetrazolium （MTT） assay and flow cytometry to observe the proliferation potential of BMSC.Results In the Western blotting analysis,positive signal lane of protein was observed in transfected group.MTT assay show that proliferation reached the peak in all groups on the fifth day,and the absorbency values of A to E group were 0.324 ± 0.027,0.319 ± 0.017,0.622 ±0.028,0.626 ± 0.020,0.778 ± 0.031.Flow cytometry show that the proliferative percentage from A to E group were 23.92 ±3.07,23.51 ±2.11,34.37 ±6.85,35.04 ± 1.45,42.56 ± 1.15.Conclusion BMP-2 or IGF-1 can promote the proliferation of BMSC under high glucose condition,but the combined has the synergy effect.

关 键 词：骨形态发生蛋白-2 胰岛素样生长因子-1 骨髓基质干细胞 高糖 细胞增殖 

分 类 号：R363[医药卫生—病理学]