Sinusoidal electromagnetic field stimulates rat osteoblast differentiation and maturation via activation of NO-cGMP-PKG pathway
Sinusoidal electromagnetic field stimulates rat osteoblast differentiation and maturation via activation of NO-cGMP-PKG pathway
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DOI：
10.1016/j.niox.2011.05.009
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发表时间：
2011-10-30
影响因子：
3.9
通讯作者：
Wang, Jiaqi
中科院分区：
文献类型：
--
作者：
Cheng, Guozheng;Zhai, Yuankun;Wang, Jiaqi
一氧化氮（NO）是重要的细胞内和细胞间信使，严重影响骨代谢。本研究旨在探讨正弦电磁场（SEMF）对成骨细胞分化成熟的影响是否通过NO-cGMP-PKG信号通路介导。我们研究了SEMF对一氧化氮合酶（NOS）活性的影响，发现一氧化氮合酶抑制剂L-NAME可以阻止SEMF介导的NOS活性和NO水平的增加。我们发现可溶性鸟苷环化酶（ODQ）的抑制剂阻断了暴露于SEMF引发的cGMP水平的增加。抑制剂PDE5可将3',5'-环gmp水解为5'-GMP，可阻止SEMFs对PKG活性的刺激。我们还阻断了NO-cGMP-PKG通路，以确定在SEMF刺激下成骨细胞的成熟和矿化是否会被抑制。通过测定碱性磷酸酶（ALP）活性、骨组织基因表达和矿化骨模量来评价。经SEMF处理后，NOS活性较对照组升高（P < 0.01），在0.5 h后达到最高水平。SEMF实验组骨组织基因表达、ALP活性和矿化骨结节均显著升高。然而，这些影响在L-NAME处理的培养物中被部分阻断。令人惊讶的是，SEMF + L-NAME组的所有成骨标志物都略高于对照培养，但低于仅暴露于SEMF的细胞。我们认为，SEMF处理激活了NO-cGMP-PKG信号通路，当该通路被阻断时，SEMF对成骨细胞分化和矿化的刺激作用减弱。(C) 2011爱思唯尔公司版权所有。
Nitric oxide (NO) is an important intracellular and intercellular messenger, critically affecting bone metabolism. The purpose of this research is to investigate whether the effect of sinusoidal electromagnetic field (SEMF) on the differentiation and maturation of osteoblasts is mediated by the NO-cGMP-PKG signal pathway. We examined the impact of SEMF on nitric oxide synthase (NOS) activity, and found that L-NAME, nitric oxide synthase's inhibitor, prevents SEMF-mediated increase in NOS activity and NO levels. We showed that an inhibitor of soluble guanylyl cyclase (ODQ) blocks the increase in cGMP levels triggered by exposure to SEMF. The inhibitor PDE5, which hydrolyzes 3',5'-cyclic-GMP to 5'-GMP, prevents the SEMFs stimulation of PKG activity. We also blocked the NO-cGMP-PKG pathway to determine whether the maturation and mineralization of osteoblasts, stimulated by SEMF, would be inhibited. This was evaluated by measuring alkaline phosphatase (ALP) activity, osterix gene expression and mineralized bone modulus. After treatment with SEMF, the NOS activity increases in comparison with the control group (P < 0.01), reaching the highest level after 0.5 h. Osterix gene expression, ALP activity and mineralized bone nodules in the SEMF experimental group also increase significantly. However, these effects are partially blocked in the L-NAME treated cultures. Surprisingly, all the osteogenic markers in the SEMF + L-NAME group were slightly higher than in the control culture, but lower than in the cells exposed to SEMF only. We conclude that the NO-cGMP-PKG signal pathway is activated by SEMF treatment, the stimulatory effect of SEMF on the differentiation and mineralization of osteoblasts is attenuated when the pathway is blocked. (C) 2011 Elsevier Inc. All rights reserved.