Xu Wenbo, Lv Shiyan, Wang Xiaoyan, Song Chengchao, Xi Chunyang, Yan Jinglong
Department of Orthopedics, The Second Affiliated Hospital of Harbin Medical University, No. 246 Xuefu Road, Harbin, 150001, Heilongjiang Province, China.
Heilongjiang Provincial Key Laboratory of Hard Tissue Development and Regeneration, Second Affiliated Hospital of Harbin Medical University, No. 246 Xuefu Road, Harbin, 150001, China.
J Orthop Surg Res. 2025 Jan 30;20(1):117. doi: 10.1186/s13018-025-05544-2.
Osteoporosis (OP) is a systemic disease characterized by low bone mass. New progress has been made in the study of OP, such as lipid peroxidation. However, the role of lipid peroxides in osteoclast differentiation is still unclear.
Bone marrow macrophages (BMMs) were extracted from C57BL/6J mice and induced to differentiate into osteoclasts, which were observed via TRAP staining, Phalloidin staining and bone pit assays. Related substances of lipid peroxidation were detected during osteoclastogenesis. The levels of osteoclastogenesis and lipid peroxides were measured by qRT-PCR, Western Blot and immunofluorescence. Activation of the p38/JNK/MAPK pathway was detected by Western Blot. The capacity for osteogenesis and angiogenesis of cells after treatment with supernatant of BMMs was evaluated. Furthermore, Ferrostatin-1 (Fer-1), from which femur and serum samples were comprehensively evaluated, was used in OVX mice.
During osteoclastogenesis, the levels of ROS, MDA, ACSL4 and LPCAT3 increased with increasing duration of RANKL stimulation, while there were no significant changes in the levels of GSH or GPX4. Fer-1 inhibited osteoclast differentiation and decreased the level of lipid peroxides. In addition, Fer-1 inhibited osteoclast-related markers by inhibiting the p38/JNK/MAPK pathway. Furthermore, the supernatant of BMMs after Fer-1 treatment promoted osteogenesis and angiogenesis. Finally, Fer-1 successfully alleviated OP in OVX mice by reducing the level of lipid peroxidation in vivo.
Fer-1 suppresses osteoclast differentiation by reducing lipid peroxidation levels regulated by ACSL4, which is mediated through the p38/JNK/MAPK signaling pathway. Additionally, Fer-1 enhances the coupling between osteogenesis and angiogenesis and has an anti-OP effect in vivo.
骨质疏松症（OP）是一种以骨量低为特征的全身性疾病。OP的研究已取得新进展，如脂质过氧化。然而，脂质过氧化物在破骨细胞分化中的作用仍不清楚。
从C57BL/6J小鼠中提取骨髓巨噬细胞（BMMs）并诱导其分化为破骨细胞，通过抗酒石酸酸性磷酸酶（TRAP）染色、鬼笔环肽染色和骨吸收陷窝测定进行观察。在破骨细胞生成过程中检测脂质过氧化的相关物质。通过实时定量聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法（Western Blot）和免疫荧光法测量破骨细胞生成和脂质过氧化物的水平。通过蛋白质免疫印迹法检测p38/应激活化蛋白激酶（JNK）/丝裂原活化蛋白激酶（MAPK）通路的激活情况。评估用BMMs上清液处理后细胞的成骨和血管生成能力。此外，在去卵巢（OVX）小鼠中使用铁死亡抑制因子1（Fer-1），并对股骨和血清样本进行全面评估。
在破骨细胞生成过程中，活性氧（ROS）、丙二醛（MDA）、长链脂酰辅酶A合成酶4（ACSL4）和溶血磷脂酰胆碱酰基转移酶3（LPCAT3）的水平随着核因子κB受体活化因子配体（RANKL）刺激时间的延长而增加，而谷胱甘肽（GSH）或谷胱甘肽过氧化物酶4（GPX4）的水平没有显著变化。Fer-1抑制破骨细胞分化并降低脂质过氧化物水平。此外，Fer-1通过抑制p38/JNK/MAPK通路抑制破骨细胞相关标志物。此外，Fer-1处理后的BMMs上清液促进成骨和血管生成。最后，Fer-1通过降低体内脂质过氧化水平成功缓解了OVX小鼠的OP。
Fer-1通过降低由ACSL4调节的脂质过氧化水平来抑制破骨细胞分化，这是通过p38/JNK/MAPK信号通路介导的。此外，Fer-1增强了成骨和血管生成之间的偶联，并在体内具有抗OP作用。
