a Department of Orthopedics , the Affiliated Hospital of Xuzhou Medical University , Xuzhou , China.
b Department of Clinical Medicine , the Affiliated Hospital of Xuzhou Medical University , Xuzhou , China.
Cell Cycle. 2018;17(17):2134-2145. doi: 10.1080/15384101.2018.1515549. Epub 2018 Sep 19.
NF-κB signaling pathway shows significant influence on wear particle-induced osteolysis, and this study aims to explore the underlying mechanism and the role of let-7f-5p in this process. A mouse calvarial osteolysis model was constructed with PMMA particles, and the bone marrow-derived macrophages (BMMs) were isolated from the osteolysis area. The expression of miRNA and protein was determined by qRT-PCR and western blot, respectively. The level of cytokines was evaluated with ELISA. Recombinant plasmids were transfected into cells for the endogenous expression of related genes. Dual-luciferase reporter assay was performed to determine the interaction between let-7f-5p and IL-10 in macrophage RAW264.7 cells. M1 macrophage polarization and expression of let-7f-5p were promoted in BMMs of osteolysis mouse model, compared with that in sham group. The expression of let-7f-5p was increased in the process of M1 macrophage polarization that induced by PMMA. Let-7f-5p was involved in M1 polarization in macrophages that treated with PMMA. IL-10 was negatively regulated by let-7f-5p. NF-κB regulated the expression of IL-10 through let-7f-5p. NF-κB participated in the PMMA-induced M1 macrophage polarization through let-7f-5p. Let-7f-5p contributed to PMMA-induced osteolysis by promoting M1 polarization of macrophages. The NF-κB/let-7f-5p/IL-10 pathway induces M1 macrophage polarization, and thus contributing to wear particle-induced osteolysis.
NF-κB 信号通路对磨屑诱导性骨溶解有显著影响，本研究旨在探讨其潜在机制及 let-7f-5p 在该过程中的作用。采用 PMMA 颗粒构建小鼠颅骨骨溶解模型，从骨溶解区分离骨髓来源的巨噬细胞（BMMs）。分别采用 qRT-PCR 和 Western blot 检测 miRNA 和蛋白的表达水平，采用 ELISA 评估细胞因子水平。转染重组质粒以细胞内源性表达相关基因。采用双荧光素酶报告基因实验检测 let-7f-5p 与巨噬细胞 RAW264.7 细胞中 IL-10 的相互作用。与 sham 组相比，骨溶解小鼠模型 BMMs 中 M1 巨噬细胞极化和 let-7f-5p 的表达增加。在 PMMA 诱导的 M1 巨噬细胞极化过程中，let-7f-5p 的表达增加。let-7f-5p 参与 PMMA 处理的巨噬细胞 M1 极化。IL-10 受 let-7f-5p 的负调控。NF-κB 通过 let-7f-5p 调节 IL-10 的表达。NF-κB 通过 let-7f-5p 参与 PMMA 诱导的 M1 巨噬细胞极化。let-7f-5p 通过促进巨噬细胞 M1 极化促进 PMMA 诱导的骨溶解。NF-κB/let-7f-5p/IL-10 通路诱导 M1 巨噬细胞极化，从而导致磨屑诱导性骨溶解。
