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  • Title: [ROLES OF SIGNAL MOLECULE p38 INVOLVED IN MINERALIZATION AND MATURATION OF OSTEOBLASTS PROMOTED BY LOW FREQUENCY PULSED ELECTROMAGNETIC FIELDS].
    Author: Fang Q, Li Z, Zhou J, Yan J, Shi W, Xie Y, Chen K.
    Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2016 Oct 08; 30(10):1238-1243. PubMed ID: 29786203.
    Abstract:
    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). CONCLUSIONS: 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. 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 mT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 mT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 mT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。.
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