Department of Pharmacy, Cardenal Herrera-CEU Universities, 46115 Valencia, Spain.
Interuniversity Research Institute for Molecular Recognition and Technological Development (IDM), Polytechnic University of Valencia, University of Valencia, 46100 Valencia, Spain.
Int J Mol Sci. 2022 Aug 1;23(15):8551. doi: 10.3390/ijms23158551.
Parathyroid hormone-related protein (PTHrP) C-terminal peptides regulate the metabolism of bone cells. PHTrP [107-111] (osteostatin) promotes bone repair in animal models of bone defects and prevents bone erosion in inflammatory arthritis. In addition to its positive effects on osteoblasts, osteostatin may inhibit bone resorption. The aim of this study was to determine the effects of osteostatin on human osteoclast differentiation and function. We used macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor κB ligand (RANKL) to induce the osteoclast differentiation of adherent human peripheral blood mononuclear cells. Tartrate-resistant acid phosphatase (TRAP) staining was performed for the detection of the osteoclasts. The function of mature osteoclasts was assessed with a pit resorption assay. Gene expression was evaluated with qRT-PCR, and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) nuclear translocation was studied by immunofluorescence. We observed that osteostatin (100, 250 and 500 nM) decreased the differentiation of osteoclasts in a concentration-dependent manner, but it did not modify the resorptive ability of mature osteoclasts. In addition, osteostatin decreased the mRNA levels of cathepsin K, osteoclast associated Ig-like receptor (OSCAR) and NFATc1. The nuclear translocation of the master transcription factor in osteoclast differentiation NFATc1 was reduced by osteostatin. Our results suggest that the anti-resorptive effects of osteostatin may be dependent on the inhibition of osteoclastogenesis. This study has shown that osteostatin controls human osteoclast differentiation in vitro through the downregulation of NFATc1.
甲状旁腺激素相关蛋白 (PTHrP) C 端肽调节骨细胞代谢。PHTrP [107-111](骨抑肽)在骨缺损动物模型中促进骨修复,并防止炎症性关节炎中的骨侵蚀。除了对成骨细胞有积极作用外,骨抑肽还可能抑制骨吸收。本研究旨在确定骨抑肽对人破骨细胞分化和功能的影响。我们使用巨噬细胞集落刺激因子 (M-CSF) 和核因子 κB 受体激活剂配体 (RANKL) 诱导贴壁人外周血单核细胞的破骨细胞分化。采用抗酒石酸酸性磷酸酶 (TRAP) 染色检测破骨细胞。通过pit 吸收测定评估成熟破骨细胞的功能。采用 qRT-PCR 评估基因表达,并用免疫荧光法研究激活 T 细胞核因子细胞质 1 (NFATc1) 的核转位。我们观察到骨抑肽 (100、250 和 500 nM) 以浓度依赖性方式降低破骨细胞的分化,但不改变成熟破骨细胞的吸收能力。此外,骨抑肽降低了组织蛋白酶 K、破骨细胞相关免疫球蛋白样受体 (OSCAR) 和 NFATc1 的 mRNA 水平。破骨细胞分化的主转录因子 NFATc1 的核转位被骨抑肽减少。我们的结果表明,骨抑肽的抗吸收作用可能依赖于对破骨细胞生成的抑制。本研究表明,骨抑肽通过下调 NFATc1 控制体外人破骨细胞分化。