脉冲电磁场促进骨髓间充质干细胞Ⅰ型胶原表达的初步研究  

A preliminary study of type Ⅰ collagen expression of bone marrow mesenchymal stem cells promoted by pulsed electromagnetic fields

作　　者：杨勇[1] 吴华[1] 陶超雄[1] 李峰[1] 李锐[1] 赵文春[2] 

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 [2]海军工程大学电力电子技术研究所

出　　处：《中华物理医学与康复杂志》2009年第7期442-445,共4页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金面上项目（50477043）

摘　　要：目的研究15Hz、1mT脉冲电磁场对骨髓间充质干细胞Ⅰ型胶原表达的影响及其机制。方法体外培养SD大鼠骨髓间充质干细胞，取第3代细胞，用15Hz、1mT脉冲电磁场每日刺激8h。用半定量逆转录聚合酶链反应（RT—PCR）检测Ⅰ型胶原mRNA的表达，用酶联免疫吸附测定（ELISA）和细胞免疫组织化学等方法检测Ⅰ型胶原的表达；加入环磷腺苷依赖的蛋白激酶A（cAMP—PKA）通路抑制剂和激动剂，并用ELISA和免疫组化的方法观察阻断和促进cAMP—PKA通路后脉冲电磁场对Ⅰ型胶原表达的影响。结果15Hz、1mT的脉冲电磁场每日刺激8h可明显促进骨髓间充质干细胞Ⅰ型胶原的表达（P〈0．05），加入cAMP—PKA通路抑制剂H-89后，脉冲电磁场促进Ⅰ型胶原表达效应明显减弱，加入cAMP—PKA通路激动剂8-Br—cAMP后脉冲电磁场促进的Ⅰ型胶原表达效应明显增强。结论15Hz、1mT脉冲电磁场可促进骨髓间充质干细胞Ⅰ型胶原的表达，脉冲电磁场促进Ⅰ型胶原表达的效应同cAMP—PKA通路密切相关。Objective To study the influence of pulsed electromagnetic fields on the expression of type Ⅰ collagen by bone marrow mesenchymal stem cells and it＇s mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro. The third passage ceils were harvested and exposed to pulsed electromagnetic fields （PEMFs） at 15 Hz and 1 mT 8 h/d for 3 days. A semi-quantitative RT-PCR technique was used to measure the type Ⅰ collagen mRNA expression; ELISA and immunohistochemistry techniques were used to measure type Ⅰ collagen expression. Inhibitors and promoters of the cAMP-dependent protein kinase A （cAMP-PKA） pathway were added. After the cAMP-PKA pathway had been inhibited or promoted, the effects of the PEMF on type Ⅰ collagen expression were measured again using ELISA and immunohistochemistry. Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type Ⅰ collagen （P≤〈0.01） in comparison with the controls. The type Ⅰ collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added, and raised when the promoter 8-Br-cAMP was added. Conclusion PEMFs at 15 Hz 1 mT can promote type Ⅰ collagen expression of bone marrow mesenchymal stem cells, and the effect is correlated with the cAMP-PKA pathway.

关 键 词：脉冲电磁场 骨髓间充质干细胞 Ⅰ型胶原 

分 类 号：R683[医药卫生—骨科学] R454.1[医药卫生—外科学]