Effect of fluoride on osteocyte-driven osteoclastic differentiation
Effect of fluoride on osteocyte-driven osteoclastic differentiation
复制标题
氟化物对骨细胞驱动的破骨细胞分化的影响。
DOI：
10.1016/j.tox.2020.152429
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发表时间：
2020-04-30
期刊：
影响因子：
4.5
通讯作者：
Xu, Hui
中科院分区：
文献类型：
--
作者：
Jiang, Ningning;Guo, Fengyang;Xu, Hui
无机氟化物的过度全身性摄取会导致骨稳态紊乱。氟骨症的发病机制仍不确定。本研究旨在研究氟化物对骨细胞驱动的破骨细胞生成的影响，并探究甲状旁腺激素（PTH）在此过程中的作用。将接种在胶原蛋白包被构建物中的IDG - SW3细胞通过矿化剂诱导分化为类骨细胞。然后，在有或无甲状旁腺激素（PTH）存在的情况下，使类骨细胞暴露于氟化物中。分别采用MTT法和蛋白质印迹法检测细胞活力以及它们产生核因子κB受体活化因子配体（RANKL）、骨保护素（OPG）和硬骨素（SOST）的能力。最后，建立一个使用接种在下层的类骨细胞和添加在上层小室中的破骨细胞前体的Transwell共培养系统，以观察在有或无PTH情况下骨细胞驱动的破骨细胞生成对氟化物的反应，并通过实时逆转录聚合酶链反应（real time RT - PCR）测量此机制中相关分子的表达。结果表明，骨细胞能耐受一定毒性剂量的氟化物，但给予PTH会显著降低骨细胞的活力。PTH放大了氟化物对骨细胞中破骨细胞生成相关分子表达的影响，但没有增强氟化物对骨细胞共培养驱动的破骨细胞生成的刺激作用。在与类骨细胞共培养且受氟化物影响的破骨细胞前体中，抗酒石酸酸性磷酸酶（TRAP）、核因子κB受体活化因子（RANK）、c - Jun氨基末端激酶（JNK）和活化T细胞核因子c1（NFAtc1）的基因表达水平显著升高。氟化物对骨细胞驱动的破骨细胞分化的影响强于PTH。总之，骨细胞在氟化物影响的破骨细胞生成的潜在机制中起着关键作用，其中涉及RANK - JNK - NFAtc1信号通路，并且PTH在此过程中有显著影响。
Excessive systemic uptake of inorganic fluorides causes disturbances of bone homeostasis. The mechanism of skeletal fluorosis is still uncertain. This study aimed to study the effect of fluoride on osteocyte-driven osteoclastogenesis and probe into the role of PTH in this process. IDG-SW3 cells seeded in collagen-coated constructs were developed into osteocyte-like cells through induction of mineral agents. Then, osteocyte-like cells were exposed to fluoride in the presence or absence of parathyroid hormone (PTH). Cell viability and their capacity to produce receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG) and sclerostin (SOST) were detected by MTT and Western blot assays, respectively. Finally, a transwell coculture system using osteocyte-like cells seeded in the low compartment, and osteoclast precursors added in the inserts was developed to observe the osteocyte-driven osteoclasogenesis response to fluoride with or without PTH, and the expression of molecules involved in this mechanism were measure by real time RT-PCR. Results showed that osteocytes withstood a toxic dose of fluoride, and yet PTH administration significantly reduced osteocytes viability. PTH amplified the effect of fluoride on the expression of osteoclastogenesis-related molecules in osteocyte, but did not enlarged the stimulating effect of fluoride on osteoclastogenesis drove by osteocyte coculture. Gene expression levels of TRAP, RANK, JNK and NFAtc1 significantly increased in fluoride affected osteoclast precursor cocultured with osteocyte-like cells. The impact of fluoride on osteocyte-driven osteoclast differentiation was stronger than that of PTH. In conclusion, osteocyte played a pivotal role on the mechanism underlying fluoride-affected osteoclastogenesis in which RANK-JNK-NFATc1 signaling pathway was involved, and PTH had a significant impact in this process.