CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

• 作者: Feng-Jie Li Jiang F Zhong Li Chen Li-Xin Xiang Ling-Yun Zou Qian Ran Shengwen Calvin Li Xiao-Mei Zhang Yan-Ni Xiao Yang Xiang Zhong-Jun Li
• 来源期刊: 世界干细胞杂志:英文版(电子版)
• 学科: 生物学
• 关键词: Irradiation Osteoporosis BONE MARROW MESENCHYMAL stem cells MONOCYTE macrophage BONE
• 年，卷（期）: 2020,（3）
• 页码范围: 222-240
• 页数: 19页
• 分类号: Q75

摘要

BACKGROUND Radiation induces rapid bone loss and enhances bone resorption and adipogenesis, leading to an increased risk of bone fracture. There is still a lack of effective preventive or therapeutic method for irradiation-induced bone injury.Receptor activator of nuclear factor κB ligand(RANKL) provides the crucial signal to induce osteoclast differentiation and plays an important role in bone resorption. However, the mechanisms of radiation-induced osteoporosis are not fully understood.AIM To investigate the role of CR6-interacting factor-1(Crif1) in osteoclastogenesis after radiation and its possible mechanism.METHODS C57 BL/6 mice were exposed to Co-60 gamma rays and received 5 Gy of wholebody sublethal irradiation at a rate of 0.69 Gy/min. For in vitro study, mouse bone marrow mesenchymal stem/stromal cells(BM-MSCs) were irradiated with Co-60 at a single dose of 9 Gy. For osteoclast induction, monocyte-macrophage RAW264.7 cells were cocultured with mouse BM-MSCs for 7 d. Clus Pro and Inter Pro Surf were used to investigate the interaction interface in Crif1 and protein kinase cyclic adenosine monophosphate(c AMP)-activited catalytic subunit alpha complex. Virtual screening using 462608 compounds from the Life Chemicals database around His120 of Crif1 was carried out using the program Autodock_vina. A tetrazolium salt(WST-8) assay was carried out to study the toxicity of compounds to different cells, including human BM-MSCs, mouse BMMSCs, and Vero cells.RESULTS Crif1 expression increased in bone marrow cells after radiation in mice.Overexpression of Crif1 in mouse BM-MSCs and radiation exposure could increase RANKL secretion and promote osteoclastogenesis in vitro. Deletion of Crif1 in BM-MSCs could reduce both adipogenesis and RANKL expression,resulting in the inhibition of osteoclastogenesis. Deletion of Crif1 in RAW264.7 cells did not affect the receptor activator of nuclear factor κB expression or osteoclast differentiation. Following treatment with protein kinase A(PKA)agonist(forskolin) and inhibitor(