中国组织工程研究2022，Vol.26Issue(23)：3691-3699,9.
白细胞介素1α 诱导破骨细胞活化和骨流失
Interleukin-1 alpha induces osteoclast activation and bone loss
摘要
Abstract
BACKGROUND: Interleukin-1 is an important pro-inflammatory cytokine that has been documented in the regulation of bone inflammation and bone remodeling. A previous study has demonstrated that interleukin-1α can induce apoptosis while inhibiting osteoblast differentiation in MC3T3-E1 cells. OBJECTIVE: To investigate the role and mechanism of interleukin-1α on osteoclast activation and bone loss in mice. METHODS: (1) Cell test: RAW264.7 cells were either treated with interleukin-1α alone or with receptor activator of nuclear factor-κB ligand (RANKL) for 1 and 4 days. Cell viability was tested by cell counting kit-8 assay. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The mRNA and protein levels of osteoclast-specific genes and genes related to nuclear factor-κB pathway and Wnt/β-catenin pathway were tested by real-time fluorescence quantitative PCR, immunofluorescence staining or western blot. Bone marrow-derived macrophages were either treated with interleukin-1α alone or with RANKL and macrophage colony-stimulating factor for 7 days. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The protein levels of osteoclast-specific genes were tested by western blot. (2) Animal test: Twenty-four male C57BL/6J mice (6-8 weeks old) were assigned into two groups at random: control group and test group. Mice were subsequently treated with interleukin-1α solution or PBS by intraperitoneal injection twice a week for 5 weeks. Bone tissues from the femurs were performed with micro-computed tomography analysis and hematoxylin-eosin staining, tartrate resistant acid phosphatase, and immunofluorescence analysis. RESULTS AND CONCLUSION: Cell test: Interleukin-1α alone significantly increased RAW264.7 cell proliferation, but stimulated cell differentiation into osteoclasts in combination with RANKL (P < 0.05). Interleukin-1α significantly increased the expression of osteoclast-related markers and the number of tartrate resistant acid phosphatase-positive multinuclear cells in RAW264.7 cells and bone marrow-derived macrophages in the existence of RANKL or RANKL+macrophage colony-stimulating factor (both P < 0.05). Interleukin-1α was found to significantly enhance the nuclear factor-κB and Wnt/beta-catenin signaling in RAW264.7 cells (P < 0.05). Blocking of nuclear factor-κB or Wnt3 signaling not only reversed the activation of nuclear factor-κB and Wnt3 signaling but also weakened the enhanced expression of osteoclast-specific genes induced by interleukin-1α in RAW264.7 cells (P < 0.05). Animal test: interleukin-1α induced bone loss in mice while also upregulating the expression of osteoclast-specific markers, RANK, TRAF6 and p65, and Wnt3 in vivo (P < 0.05). The findings indicate that interleukin-1α can induce osteoclast activation and bone loss by promoting the nuclear factor-κB and Wnt signaling pathways.关键词
白细胞介素1 α/破骨细胞/RAW264.7细胞/骨髓来源巨噬细胞/抗酒石酸酸性磷酸酶/核因子κB受体活化因子配体/骨丢失/μCT/核转录因子κB信号通路/Wnt/β-catenin信号通路Key words
interleukin-1α/osteoclast/RAW264.7 cell/bone marrow-derived macrophage/tartrate resistant acid /phosphatase/receptor activator of nuclear factor-κB ligand/bone loss/micro-computed tomography/nuclear factor-κB signaling/Wnt/β-catenin signaling分类
医药卫生引用本文复制引用
杨锐娟,李阳阳,蔡瑞艳,刘慧兵,郭春..白细胞介素1α 诱导破骨细胞活化和骨流失[J].中国组织工程研究,2022,26(23):3691-3699,9.基金项目
the National Natural Science Foundation of China,No.81672190(to GC) （to GC）
the Project of Health Commission of Henan Province,No.SB201903013(to GC) （to GC）
the Project of Science and Technology Department of Henan Province,No.182102310260(to GC) （to GC）
the Youth Foundation of the First Affiliated Hospital of Xinxiang Medical University,No.QN-2020-B11(to YRJ)基金资助:国家自然科学基金资助项目(81672190),项目负责人:郭春 （to YRJ）
河南省卫生健康委员会项目(SB201903013),项目负责人:郭春 （SB201903013）
河南省科学技术厅项目(182102310260),项目负责人:郭春 （182102310260）
新乡医学院第一附属医院青年基金项目(QN-2020-B11),项目负责人:杨锐娟 （QN-2020-B11）
