4.7 Article

The interdependent relationship between the nitric oxide signaling pathway and primary cilia in pulse electromagnetic field-stimulated osteoblastic differentiation

期刊

FASEB JOURNAL
卷 36, 期 6, 页码 -

出版社

WILEY
DOI: 10.1096/fj.202101577RR

关键词

endothelial nitric oxide synthase; nitric oxide; osteogenic differentiation; primary cilia; pulsed electromagnetic fields

资金

  1. National Natural Science Foundation of China [81270963, 81770879]

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This study found that pulsed electromagnetic fields (PEMFs) can promote osteoblastic differentiation by activating the NOS/NO/sGC/cGMP/PKG signaling pathway, which requires the presence of primary cilia. The study also observed the activation of the NO/cGMP signaling pathway in the ciliary compartment and showed that PEMFs cannot stimulate osteoblastic differentiation if the NO signaling pathway is blocked or ciliogenesis is inhibited.
Pulsed electromagnetic fields (PEMFs) have long been recognized being safe and effective in treating bone fracture nonunion and osteoporosis. However, the mechanism of osteogenic action of PEMFs is still unclear. While primary cilia are reported to be a sensory organelle for PEMFs, and nitric oxide (NO) plays an indispensable role in osteogenic effect of PEMFs, the relationship between NO and primary cilia is unknown. In this study, effects of treatment with 50 Hz 0.6 mT PEMFs on osteogenic differentiation and mineralization, NO secretion, and ciliary location of specific proteins were examined in rat calvarial osteoblasts (ROBs) with normal or abrogated primary cilia. It was found that PEMFs stimulated the osteogenic differentiation by activating the NOS/NO/sGC/cGMP/PKG signaling pathway, which need the existence of primary cilia. All components of the signaling pathway including iNOS, eNOS, sGC, PKG-1, and PKG-2 were localized to primary cilia, and eNOS was phosphorylated inside the primary cilia. Besides, primary cilia were elongated significantly by PEMF treatment and changed dynamically with the activation NO/cGMP pathway. When the pathway was blocked by L-NAME, PEMFs could no longer elongate the primary cilia and stimulate the osteoblastic differentiation. Thus, this study for the first time observed activation of the NO/cGMP signaling pathway in ciliary compartment of osteoblasts, and PEMFs could not stimulate the osteoblastic differentiation if the NO signaling pathway was blocked or the ciliogenesis was inhibited. Our findings indicate the interdependent relationship between NO and primary cilia in the PEMF-promoted osteogenesis.

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