[低频脉冲电磁场诱导大鼠骨髓间充质干细胞中ACTN2、α-肌动蛋白和TNNT2的表达]

[Expression of ACTN2, alpha-actin and TNNT2 in rat bone marrow-derived mesenchymal stem cells induced by low frequency pulsed electromagnetic fields].

作者信息

Li Kai, Jiang Jian, Yao Xiao-Lin, Qiu Fei-Yuan, Yan Zhong-Qin, Wu Wen-Chao, Liu Xiao-Jing, Li Liang

机构信息

Institute of Biomedical Engineering, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2012 Sep;43(5):670-4, 710.

PMID:23230735

Abstract

OBJECTIVE

To study the expression of Actinin, alpha 2 (ACTN2), alpha-actin and Troponin T type 2 (TNNT2) in rat bone marrow-derived mesenchymal stem cells (rBMSCs) induced by low frequency pulsed electromagnetic fields (PEMFs).

METHODS

The rBMSCs were isolated by adherence method and the third-generation of the rBMSCs were randomly divided into PEMFs groups, 5-Azacytidine groups (5-Aza), PEMFs +5-Aza group and control groups. PEMFs groups with complete medium were exposed to 50 Hz, 1 mT PEMFs for 30 min every day, lasting for 10 d, 15 d and 20 d, respectively. 5-Aza groups were induced by 10 micromol/L 5-Aza for 1 day, then the medium was changed to complete medium without 5-Aza. PEMFs +5-Aza group were firstly induced by 10 micromol/L 5-Aza for 1 day, and then were exposed to PEMFs according to the way of PEMFs group. And control groups were only cultured with complete medium, rBMSCs' growth status and morphological features were observed by inverted phase microscope. The mRNA expressions of ACTN2 and TNNT2 were determined by real-time fluorescent quantitation PCR. And the protein expression of the ACTN2, alpha-actin and TNNT2 were detected with Western blot method.

RESULTS

The result showed that the mRNA expression level of the ACTN2 and TNNT2 and the protein expression level of the ACTN2, alpha-actin and TNNT2 were significantly higher in the PEMFs group, 5-Aza group and PEMFs+ 5-Aza group when compared with the control group.

CONCLUSION

PEMFs may induce rBMSCs differentiation into cardiomyocyte-like cells in vitro.

摘要

目的

研究低频脉冲电磁场（PEMFs）诱导大鼠骨髓间充质干细胞（rBMSCs）中辅肌动蛋白α2（ACTN2）、α-肌动蛋白和肌钙蛋白T2型（TNNT2）的表达。

方法

采用贴壁法分离rBMSCs，将第三代rBMSCs随机分为PEMFs组、5-氮杂胞苷组（5-Aza）、PEMFs + 5-Aza组和对照组。PEMFs组用完全培养基，每天暴露于50 Hz、1 mT的PEMFs 30分钟，分别持续10天、15天和20天。5-Aza组用10 μmol/L 5-Aza诱导1天，然后更换为不含5-Aza的完全培养基。PEMFs + 5-Aza组先用10 μmol/L 5-Aza诱导1天，然后按PEMFs组的方式暴露于PEMFs。对照组仅用完全培养基培养，通过倒置相差显微镜观察rBMSCs的生长状态和形态特征。采用实时荧光定量PCR测定ACTN2和TNNT2的mRNA表达。并用蛋白质印迹法检测ACTN2、α-肌动蛋白和TNNT2的蛋白表达。