OBJECTIVE: To examine the effect of JMJD1C on the osteogenesis of BMSCs and the potential underlying mechanism.
METHODS: BMSCs were isolated from mouse bone marrow tissues. Oil Red O staining, Alizarin red staining, alkaline phosphatase staining and the expression of adipogenic and osteogenic-associated genes were assessed to determine the differentiation of BMSCs. Bone marrow-derived macrophages (BMMs) were incubated with receptor activator of nuclear factor-kappa Β ligand to induce osteoclast differentiation, and osteoclast differentiation was confirmed by tartrate-resistant acid phosphatase staining. Other related genes were measured via reverse transcription coupled to the quantitative polymerase chain reaction and western blotting. Enzyme-linked immunosorbent assays were used to measure the levels of inflammatory cytokines, including tumor necrosis factor alpha, interleukin-6 and interleukin-1 beta.
RESULTS: The osteogenic and adipogenic differentiation potential of BMSCs isolated from mouse bone marrow samples was evaluated. JMJD1C mRNA and protein expression was upregulated in BMSCs after osteoblast induction, while p-nuclear factor-κB (NF-κB) and inflammatory cytokines were not significantly altered. Knockdown of JMJD1C repressed osteogenic differentiation and enhanced NF-κB activation and inflammatory cytokine release in BMSCs. Moreover, JMJD1C expression decreased during BMM osteoclast differentiation.
CONCLUSIONS: The JMJD1C/NF-κB signaling pathway is potentially involved in BMSC osteogenic differentiation and may play vital roles in the pathogenesis of osteoporosis.
目的:研究JMJD1C对骨髓间充质干细胞成骨的影响及其潜在机制。
方法:从小鼠骨髓组织中分离BMSCs。油红O染色,茜素红染色,碱性磷酸酶染色和成脂和成骨相关基因的表达进行评估以确定BMSCs的分化。将骨髓源性巨噬细胞(BMMs)与核因子κB配体的受体激活剂孵育以诱导破骨细胞分化,抗酒石酸酸性磷酸酶染色证实了破骨细胞的分化。通过与定量聚合酶链反应和蛋白质印迹偶联的逆转录来测量其他相关基因。酶联免疫吸附试验用于测量炎症细胞因子的水平,包括肿瘤坏死因子α,白细胞介素-6和白细胞介素-1β。
结果:评价了从小鼠骨髓样品中分离的BMSCs的成骨和成脂分化潜能。JMJD1C在BMSCs诱导成骨细胞后mRNA和蛋白表达上调,而P-核因子-κB(NF-κB)和炎性细胞因子无明显改变。敲除JMJD1C抑制BMSCs成骨分化并增强NF-κB活化和炎性细胞因子释放。此外,在BMM破骨细胞分化过程中JMJD1C表达降低。
结论:JMJD1C/NF-κB信号通路可能参与BMSC的成骨分化,并可能在骨质疏松的发病中起重要作用。