雷帕霉素与低频电磁场联合应用对大鼠骨髓间充质干细胞成骨效应的影响  

The synergistic effect of rapamycin and a low frequency pulsed electromagnetic field on the genesis of bone marrow mesenchymal stem cells

作　　者：易智谦[1] 宋明宇[1] 虞冀哲[1] 刘阳[1] 韦盛[1] 杨勇[1] 李峰[1] 吴华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《中华物理医学与康复杂志》2013年第5期342-347,共6页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金资助项目（51077065）;湖北省自然科学基金（2011CDC032）

摘　　要：目的观察雷帕霉素与低频电磁场联合运用对大鼠骨髓间充质干细胞（BMSCs）成骨效应的影响。方法选取成年SD大鼠15只，采用体外分离法培养大鼠BMSCs，传代3次后接种至细胞培养板中，将其分为对照组、雷帕霉素组、电磁场刺激组和联合应用组。对照组应用达尔伯改良伊格尔培养基（DMEM）进行培养；雷帕霉素组在DMEM中加入浓度为5nM的雷帕霉素；电磁场刺激组则在DMEM基础上给予频率为15Hz、强度为1mT的脉冲电磁场刺激；联合应用组在加入浓度为5nM雷帕霉素的DMEM基础上给予频率为15Hz、强度为1mT，每天1h的脉冲电磁场刺激。分别于干预后7d、14d和21d，提取细胞的总RNA和总蛋白，采用茜素红染色后置于显微镜下观察，分析比较不同时间点各组的成骨效应。Objective To study whether Rapamycin and a low frequency pulsed electromagnetic field （LFPEMF） have synergistic effects on osteogenesis. Methods Bone marrow mesenchymal stem cells （BMSCs） were isolated from 4-week-old Sprague-Dawley rats （male, 60-80 g） and cultured. The passage three BMSCs were transplanted into six well plates. The wells were divided into a control group in which the ceils were cultivated in Dul- becco＇s modification of Eagle＇s medium （DMEM） , a rapamycin group in which the cells were cultivated in DMEM containing 5 nM rapamyein, an electromagnetic field group in which the cells were cultivated in DMEM in a LFPEMF, and a combined treatment group in which the cells were cultivated in DMEM containing rapamycin in a LFPEMF. Proteins and RNA were extracted and the calcium disposition was stained using alizarin red after 7, 14 and 21 days of incubation. Real-time PCR or western blotting was then used to detect any differences in the expression of the RUNX2, BSP, OPN or BMP2 genes.

关 键 词：雷帕霉素 成骨诱导剂 电磁场 联合成骨 

分 类 号：R[医药卫生]