信号分子p38参与低频脉冲电磁场促进成骨细胞矿化成熟的实验研究  

ROLES OF SIGNAL MOLECULE p38 INVOLVED IN MINERALIZATION AND MATURATION OF OSTEOBLASTS PROMOTED BY LOW FREQUENCY PULSED ELECTROMAGNETIC FIELDS

作　　者：方清清 李志忠[2] 周建[1] 闫娟丽[1] 石文贵[1] 谢艳芳[1] 陈克明[1] 

机构地区：[1]兰州军区兰州总医院全军创伤骨科研究所,兰州730050 [2]兰州理工大学生命科学与工程学院

出　　处：《中国修复重建外科杂志》2016年第10期1238-1243,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81270963;81471090)~~

摘　　要：目的研究信号分子丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)是否参与50 Hz、0.6 m T低频脉冲电磁场促进大鼠颅骨成骨细胞矿化成熟的过程。方法取出生48 h内的SPF级Wistar大鼠6只,采用酶消化法分离培养大鼠颅骨成骨细胞。取原代颅骨成骨细胞,经50 Hz、0.6 m T低频脉冲电磁场处理0、5、10、20、40、60、120 min后,采用Western blot检测MAPKs的磷酸化水平。将成骨细胞随机分为对照组(A组)、低频脉冲电磁场处理组(B组)、p-p38抑制剂SB202190组(C组)、SB202190+低频脉冲电磁场处理组(D组),经相应处理1、4、7 d后检测其ALP活性。将90%以上融合的成骨细胞经成骨诱导处理后同上法分组,处理9 d时行ALP组织化学染色,12 d时行茜素红染色观察。结果 Western blot检测示,处理各时间点磷酸化细胞外信号调节激酶1/2、磷酸化c-Jun氨基末端激酶1/2蛋白表达量无显著变化(P>0.05);而p-p38蛋白表达量则在处理5 min后开始升高,至40 min时达峰值,之后开始逐步下降,但各时间点均显著高于0 min时(P<0.05)。B组ALP活性、ALP阳性克隆数、ALP阳性克隆面积、钙结节数及钙结节面积均显著多于A、C、D组(P<0.05)。结论在50 Hz、0.6 m T低频脉冲电磁场促进大鼠颅骨成骨细胞矿化成熟的过程中,信号分子p38参与其中。Objective To investigate whether signal molecule mitogen-activated protein kinases （MAPKs） involves in the process of the mineralization and maturation of rat calvarial osteoblasts promoted by 50 Hz, 0.6 mT pulsed electromagnetic fields. Methods Rat calvarial osteoblasts were obtained by enzyme digestion from the skull of 6 neonatal Wistar rats of SPF level. The primary osteoblasts were treated in 50 Hz and 0.6 mT pulsed electromagnetic fields for 0, 5, 10, 20, 40, 60, and 120 minutes; the protein expression of phosphorylated MAPKs was detected by Western blot. The osteoblasts were randomly divided into group A （control group）, group B （low frequency pulse electromagnetic fields treatment group）, group C （SB202190 group）, and group D （SB202190＋low frequency pulse electromagnetic fields treatment group）; the alkaline phosphatase （ALP） activities were tested after corresponding treatment for 1, 4, and 7 days. The corresponding treated more than 90% confluenced osteoblasts were cultured under condition of osteogenic induction, then ALP staining and alizarin red staining were carried out at 9 and 12 days respectively. Results The results of Western blot showed that there was no significant changes in the protein expressions of phosphorylated level of extracellular signal-related kinases 1/2 and c-Jun amino N-terminal kinases 1/2 in 50 Hz, 0.6 mT pulsed electromagnetic fields （P〉0.05）, but the phosphorylated level of p38 began to increase at 5 minutes, peaked at 40 minutes, then gradually decreased, and it was significantly higher at 5-120 minutes than at 0 minute （P〈0.05）. After the activities of p-p38 was inhibited by inhibitor SB202190, the ALP activities, positive colonies and area of ALP and calcified nodules of group B were significantly higher than groups A, C, and D （P〈0.05）. Conclusion p38 is one of the signal molecules involved in the process of the mineralization and maturation of rat calvarial osteoblasts promoted by 50 Hz, 0.6 mT pulsed electromagnetic fields.

关 键 词：骨质疏松症 脉冲电磁场 成骨细胞 丝裂原活化蛋白激酶 

分 类 号：R580[医药卫生—内分泌]