体外小儿骨髓间充质干细胞对牵张应力早期应答反应的研究  

Early responses of bone marrow mesenchymal stem cells to mechanical strain in children

作　　者：黎润光[1] 邵景范[1] 魏明发[1] 洪振亚[2] 谢柏臻[3] 杨小进[1] 张文[1] 陈超[1] 王子民[1] 袁继炎[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院小儿外科,武汉430030 [2]华中科技大学同济医学院附属同济医院血液科,武汉430030 [3]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《中华小儿外科杂志》2008年第5期294-297,共4页Chinese Journal of Pediatric Surgery

基　　金：国家教委留学回国人员启动基金（编号：013456）

摘　　要：目的观察体外小儿骨髓间充质干细胞（MSCs）对周期性牵张应力的早期应答反应。方法分离并培养小儿MSCs，利用体外细胞牵张应力装置对第三一六代细胞加载短期力学信号，检测各组细胞内Ca^2＋以及早期反应基因蛋白（c-fos、c-jun和c-myc）的表达情况。结果应力干预MSCs后，细胞内Ca^2＋浓度在受力1min后即增高，荧光强度是对照组的164．63％，3、5min升高更明显，干预10min后Ca^2＋浓度升高的幅度有下降趋势；c-los、mjun和c-myc基因蛋白在未受力组细胞内不表达或微量表达，加载力学信号后，表达发生了明显的变化，随时间的延长，均表现为增强-持续-回复原来水平。结论周期性牵张应力影响MSCs内Ca^2＋的浓度及早期反应基因蛋白（c-los、mjun和mmyc）的表达，推测c-los、c-jun和c-myc参与了MSCs响应力学的信号传导途径，可能依赖Ca^2＋通道的活化来调控。Objective To study the early responses of bone marrow mesenchymal stem cells （MSCs）to mechanical strain in childrerL Methods The MSCs were isolated from children and cultured primarily. After 3 to 6 generations, the MSCs were stretched mechanically. Intracellular free [Ca^2＋ ] was measured with flow cytometry. The early-response genes/proteins （c-los, c-jun, c-myc）were examined with RT-PCR,immunocytochemistry and Western Blot. Results Intracellular free [Ca^2＋ ]i was elevated after one minute of mechanical stretch. The fluorescence intensity was 164. 63% that of the control group. The increase was more prominent at three and five minutes. It became less obvious at 10 minutes. The application of mechanical strain also increased intracellular c-los, c-jun, c-myc mRNA / protein levels. Conclusions These data suggest mechanical strain could affect MSCs [Ca^2＋ ]i and early- response genes/proteins levels of c-fos,c-jun,c-myc,implying that they are involved in the early mechanical strain transduction pathways,possibly by activating calcium ion channel in MSCs.

关 键 词：间质干细胞 牵张应力 钙通道 

分 类 号：R72[医药卫生—儿科]