Liu Ya Dong, Liu Jian Feng, Liu Bin
Department of Spine Surgery, The First Hospital of Jilin University, Changchun, China.
Department of Hand Surgery, The First Hospital of Jilin University, Changchun, China.
Bone Joint Res. 2022 Apr;11(4):200-209. doi: 10.1302/2046-3758.114.BJR-2020-0308.R2.
The role of N,N-dimethylformamide (DMF) in diabetes-induced osteoporosis (DM-OS) progression remains unclear. Here, we aimed to explore the effect of DMF on DM-OS development.
Diabetic models of mice, RAW 264.7 cells, and bone marrow macrophages (BMMs) were established by streptozotocin stimulation, high glucose treatment, and receptor activator of nuclear factor-κB ligand (RANKL) treatment, respectively. The effects of DMF on DM-OS development in these models were examined by micro-CT analysis, haematoxylin and eosin (H&E) staining, osteoclast differentiation of RAW 264.7 cells and BMMs, H&E and tartrate-resistant acid phosphatase (TRAP) staining, enzyme-linked immunosorbent assay (ELISA) of TRAP5b and c-terminal telopeptides of type 1 (CTX1) analyses, reactive oxygen species (ROS) analysis, quantitative reverse transcription polymerase chain reaction (qRT-PCR), Cell Counting Kit-8 (CCK-8) assay, and Western blot.
The established diabetic mice were more sensitive to ovariectomy (OVX)-induced osteoporosis, and DMF treatment inhibited the sensitivity. OVX-treated diabetic mice exhibited higher TRAP5b and c-terminal telopeptides of type 1 (CTX1) levels, and DMF treatment inhibited the enhancement. DMF reduced RAW 264.7 cell viability. Glucose treatment enhanced the levels of TRAP5b, cathepsin K, Atp6v0d2, and H-ATPase, ROS, while DMF reversed this phenotype. The glucose-increased protein levels were inhibited by DMF in cells treated with RANKL. The expression levels of antioxidant enzymes Gclc, Gclm, Ho-1, and Nqo1 were upregulated by DMF. DMF attenuated high glucose-caused osteoclast differentiation by targeting mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signalling in BMMs.
DMF inhibits high glucose-induced osteoporosis by targeting MAPK and NF-κB signalling. Cite this article: 2022;11(4):200-209.
N，N-二甲基甲酰胺（DMF）在糖尿病性骨质疏松症（DM-OS）进展中的作用尚不清楚。在此，我们旨在探讨DMF对DM-OS发展的影响。
分别通过链脲佐菌素刺激、高糖处理和核因子κB受体活化因子配体（RANKL）处理建立小鼠糖尿病模型、RAW 264.7细胞模型和骨髓巨噬细胞（BMM）模型。通过显微CT分析、苏木精和伊红（H&E）染色、RAW 264.7细胞和BMM的破骨细胞分化、H&E和抗酒石酸酸性磷酸酶（TRAP）染色、TRAP5b和I型胶原C端肽（CTX1）的酶联免疫吸附测定（ELISA）分析、活性氧（ROS）分析、定量逆转录聚合酶链反应（qRT-PCR）、细胞计数试剂盒-8（CCK-8）测定和蛋白质免疫印迹法检测DMF对这些模型中DM-OS发展的影响。
建立的糖尿病小鼠对卵巢切除（OVX）诱导的骨质疏松更敏感，而DMF治疗可抑制这种敏感性。接受OVX治疗的糖尿病小鼠表现出较高的TRAP5b和I型胶原C端肽（CTX1）水平，而DMF治疗可抑制这种升高。DMF降低了RAW 264.7细胞的活力。高糖处理提高了TRAP5b、组织蛋白酶K、Atp6v0d2和H-ATPase、ROS的水平，而DMF逆转了这种表型。在RANKL处理的细胞中，DMF抑制了高糖增加的蛋白质水平。DMF上调了抗氧化酶Gclc、Gclm、Ho-1和Nqo1的表达水平。DMF通过靶向BMM中的丝裂原活化蛋白激酶（MAPK）和核因子κB（NF-κB）信号通路减弱了高糖诱导的破骨细胞分化。
DMF通过靶向MAPK和NF-κB信号通路抑制高糖诱导的骨质疏松。引用本文：2022；11（4）：200-209。
