中国骨质疏松杂志

远端上游元件结合蛋白3对成骨细胞增殖、成骨分化和糖酵解的影响

Effects of far upstream element-binding protein 3 on osteoblast proliferation, osteogenic differentiation, and glycolysis

DOI：10.3969/j.issn.1006-7108.2024.07.010

中文关键词: 远端上游元件结合蛋白3 糖酵解成骨细胞增殖成骨分化骨质疏松性骨折

英文关键词:far upstream element binding protein3 glycolysis osteoblast proliferation osteogenic differentiation osteoporosis fracture

基金项目:2023年度中山市医学科研项目（20231A020120）

作者	单位
姚尚辰1 赵波1 李艳清1 李林2*	1.中山市骨科医院骨科，广东中山 528400 2.延边大学附属医院骨科，吉林延边 133000

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中文摘要:

目的探究远端上游元件结合蛋白3（far upstream element binding protein 3，FUBP3）介导糖酵解对成骨细胞分化能力和功能的影响。方法实时荧光定量PCR（reverse transcription real-time fluorescence quantitative PCR，RT-qPCR）检测骨质疏松性骨折患者外周血和接受入院24 h、接受治疗1、2、3、4周时FUBP3 mRNA的表达水平，RT-qPCR和蛋白免疫印迹法（Western blot）检测骨质疏松性骨折患者骨组织FUBP3 mRNA和蛋白表达水平。MC3T3-E1细胞分为sh-NC组、sh-FUBP3组、pcDNA组、pcDNA-FUBP3组，MTT和EdU检测细胞增殖活性，Western blot检测Runt相关转录因子2（runtrelated transcription factor 2，Runx2）、骨桥蛋白（osteopontin，OPN）、葡萄糖转运蛋白1（glucose transporters type 1，Glut1）、Glut3蛋白，试剂盒检测碱性磷酸酶（alkaline phosphatse，ALP）活性、葡萄糖消耗量及乳酸生成量。结果骨质疏松性骨折患者外周血中FUBP3 mRNA表达水平高于非骨质疏松性骨折患者，骨质疏松性骨折患者骨组织中FUBP3 mRNA和蛋白表达水平高于非骨质疏松性骨折患者（P＜0.05）；骨质疏松性骨折患者在入院24 h、接受治疗1、2、3、4周时外周血，发现随着治疗时间增加FUBP3 mRNA逐渐降低，呈时间依赖性，在治疗4周时表达水平最低（P＜0.05）。与sh-NC组相比，sh-FUBP3组FUBP3 mRNA和蛋白表达明显降低，细胞活性和EdU阳性细胞比率、葡萄糖消耗量、乳酸生成量、ALP活性明显增加，Glut1、Glut3、Runx2、OPN蛋白表达上调（P＜0.05）；与pcDNA组相比，pcDNA-FUBP3组FUBP3 mRNA和蛋白表达明显增加，细胞活性和EdU阳性细胞比率、葡萄糖消耗量、乳酸生成量、ALP活性明显降低，Glut1、Glut3、Runx2、OPN蛋白表达下调（P＜0.05）。结论骨质疏松性骨折患者外周血和骨组织中FUBP3表达上调，敲低FUBP3抑制成骨细胞增殖、成骨分化和糖酵解，过表达FUBP3则相反。

英文摘要:

Objective To investigate the effect of far upstream element binding protein 3 (FUBP3) mediated glycolysis on the differentiation and function of osteoblasts. Methods RT-qPCR was used to detect FUBP3 mRNA expression in peripheral blood of patients with brittleness fracture at 24 h after admission, and 1, 2, 3, and 4 w after treatment. RT-qPCR and Western blotting were used to detect FUBP3 mRNA and protein expression in the bone tissue of patients with fragility fracture. MC3T3-E1 cells were divided into sh-NC group, sh-FUBP3 group, pcDNA group, and PCDNA-FUBP3 group. The cell proliferation activity was detected with MTT and EdU. Western blotting was used to detect Runt-related transcription factor 2 (Runx2), osteopontin (OPN), glucose transporter 1(Glut1), and Glut3 proteins. Alkaline phosphatase (ALP) activity, glucose consumption, and lactic acid production were measured. Results The expression level of FUBP3 mRNA in peripheral blood of patients with fragility fracture was higher than that of patients with non-fragility fracture, and the expression levels of FUBP3 mRNA and protein in the bone tissue of patients with fragility fracture were higher than those in patients with non-fragility fracture (P<0.05). In peripheral blood of fragility fracture patients at 24 h after admission and 1, 2, 3 and 4 w after treatment, FUBP3 mRNA was found to decrease gradually with the increase of treatment time in a time-dependent manner. The expression level was the lowest at 4 w after treatment (P<0.05). Compared to those in sh-NC group, the mRNA and protein expressions of FUBP3 in sh-FUBP3 group decreased significantly. The cell activity, EdU positive cell ratio, glucose consumption, lactic acid production, and ALP activity increased significantly. The protein expressions of Glut1, Glut3, Runx2 and OPN were up-regulated (P<0.05). Compared to those in pcDNA group, the expressions of FUBP3 mRNA and protein in PCDNA-FUBP3 group increased significantly, while the cell activity, EdU positive cell ratio, glucose consumption, lactic acid production and ALP activity decreased significantly. The protein expressions of Glut1, Glut3, Runx2 and OPN were down-regulated (P<0.05). Conclusion FUBP3 expression is up-regulated in peripheral blood and bone tissue of patients with osteoporotic fractures. Knockdown of FUBP3 inhibits osteoblast proliferation, osteogenic differentiation, and glycolysis. Overexpression of FUBP3 inhibits the opposite.

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