【摘要】 目的:探究己糖激酶2(hexokinase 2,HK2)调控根尖周炎骨吸收的分子机制。方法:通过RNA测序分析人根尖周囊肿、肉芽肿与正常黏膜的糖酵解基因表达差异；免疫组织化学法检测HK2蛋白表达。体外实验：骨髓来源的巨噬细胞(bone marrow-derived macrophages, BMDMs)经脂多糖(lipopolysaccharide, LPS)和己糖激酶抑制剂2-脱氧-D-葡萄糖(2-deoxy-D-glucose, 2DG)处理后，采用Western blot检测干扰素基因刺激蛋白(stimulator of interferon genes, STING)通路激活情况，抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase, TRAP)染色评估破骨分化能力。体内实验：建立C57/BL6小鼠根尖周炎模型并对小鼠腹腔注射2DG(14 d/28 d),通过micro-CT量化骨吸收情况，根尖周炎组织切片TRAP染色计数破骨细胞，免疫组织化学检测HK2和STING表达。结果:人根尖周炎组织糖酵解基因显著上调，免疫组化结果显示HK2表达明显增高。2DG抑制HK2后，LPS诱导的BMDMs细胞STING表达降低23.6%(P<0.05),破骨分化减少22.2%(P<0.01)。动物实验显示2DG干预小鼠根尖周炎骨吸收减少，破骨细胞数量下降，STING表达降低。结论:HK2介导的糖酵解通过激活STING通路促进巨噬细胞破骨分化，抑制HK2可减轻根尖周炎骨吸收。更多还原

【Abstract】 Objective: To investigate the molecular mechanism by which hexokinase 2(HK2) regulates bone resorption in apical periodontitis. Methods: Using RNA sequencing, the differences in glycolysis gene expression between human apical cysts/granulomas and normal mucosa were analyzed. Protein expression of HK2 was detected by immunohistochemistry. In vitro experiments: bone marrow-derived macrophages(BMDMs) were treated with lipopolysaccharides(LPS) and HK2 inhibitor 2-deoxy-D-glucose(2DG). Activation of the stimulator of interferon genes(STING) pathway was analyzed by Western blot, and osteoclast differentiation ability was assessed by tartrate resistant acid phosphatase(TRAP) staining. In vivo experiments: C57/BL6 mice were intraperitoneally injected with 2DG while establishing a periapical periodontitis model(14 days/28 days). Bone resorption was quantified by micro-CT, osteoclasts were counted by TRAP staining in apical periodontitis tissue sections, and the expressions of HK2 and STING were detected by immunohistochemistry. Results: Glycolysis-related genes were significantly up-regulated in apical periodontitis tissues. Immunohistochemical results showed that the expression of HK2 was significantly increased. After inhibition of HK2 by 2DG, the expression level of STING in LPS-induced BMDMs was decreased by 23.6%(P<0.05), and osteoclast differentiation induced by LPS in BMDMs was reduced by 22.2%(P<0.01). Animal experiments showed that 2DG treatment reduced the bone resorption, osteoclast number, and STING expression in mice with apical periodontitis. Conclusion: HK2-mediated glycolysis promotes macrophage osteoclastic differentiation by activating the STING signaling pathway. Inhibition of HK2 can alleviate bone resorption in apical periodontitis.更多还原