电磁场对骨髓间充质干细胞聚集蛋白聚糖I、II型胶原和Sox9等基因表达的影响  

Effects of electromagnetic fields on the expressions of aggrecan I and II collagen and Sox9 by bone marrow mesenchymal stem cells

作　　者：刘宇[1,2] 吴华[1] 刘朝旭[1] 李贵振[1] 赵文春[3] 杨勇[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 [2]湖北省十堰市郧阳医学院附属东风医院骨科 [3]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8期519-523,共5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：目的探讨15Hz、1mT电磁场（EMFs）对骨髓间充质干细胞成软骨分化指标聚集蛋白聚糖（Agc）I、II型胶原和Sox9等的影响及其机制。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、1mTEMFs刺激，8h／d。采用半定量逆转录聚合酶链反应（RT—PCR）法检测甲状旁腺激素受体相关肽（PTHrp）、AgcI、II型胶原和Sox9等mRNA的表达，用Western Blot检测II型胶原蛋白的表达。加入解蛋白激酶A（PKA）抑制剂H－89和蛋白激酶C（PKC）抑制剂Go－6976，并再次用RT—PCR法和Western Blot观察EMFs对AgcI、II型胶原和Sox9等mRNA的表达的影响。结果EMFs促进骨髓间充质干细胞PTHrp、AgcI、II型胶原和Sox9等mRNA的表达，促进II型胶原蛋白的表达；在加入H－89（10μm）和Go－6976（12μm）后EMFs的这种促进AgcI、II型胶原的效应就明显减弱，但Sox9的表达不受影响。结论EMFs促进骨髓间充质干细胞成软骨分化指标AgcI、II型胶原和Sox9等mRNA的表达，这种促进作用同PKA和PKC通路有联系。Objective To study the expressions of aggrecan （Age） I and II collagen and Sox9 by bone marrow mesenehymal stem cells exposed to electromagnetic fields （EMFs） and it＇s mechanisms involved. Methods Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and cultured in vitro. The third passage cells were harvested and exposed to 15 Hz 1 mT EMFs for 8 h/d. The semi-quantitative reverse transeription-polymerase chain reaction （RT-PCR） technique was used to measure parathyroid hormon receptor related peptide （PTHrp） , Age I and Ⅱ collagen and Sox9 mRNA. Western blotting was used to measure type II collagen expression. After the inhibitor of protein kinase A （PKA） H-89 and the inhibitor of protein kinase C （PKC） Go-6976 （ 12 μm） were added, the effects of EMFs on Age I and II collagen and Sox9 mRNA expressions were measured again by using RT- PCR, and Western blotting technique. Results The EMFs induced significant increase of mRNA expressions of PTHrp, Age I and II collagen and Sox9 in comparison to the controls,and promoted type II collagen protein expression. The Age I and II collagen expressions decreased after PKA pathway inhibitor H-89 and PKC inhibitor Go-6976 were added, but the mRNA expression of Sox9 was not affected. Conclusion This study shows 15Hz lmT EMFs can promote mRNA expressions of Age I and II collagan and Sox9 of ehondrogenesis differentiation markers in bone marrow mesenchymal stem cells. The effect is correlated with PKA and PKC pathways.

关 键 词：电磁场 骨髓间充质干细胞 聚集蛋白聚糖I型胶原 聚集蛋白聚糖II型胶原 

分 类 号：R68[医药卫生—骨科学] R454.1[医药卫生—外科学]