口腔疾病防治 ›› 2020, Vol. 28 ›› Issue (1): 24-29.DOI: 10.12016/j.issn.2096-1456.2020.01.004

• 基础研究 • 上一篇    下一篇

FoxO1在1,25(OH)2D3调控高糖环境下成骨细胞代谢中的作用

周佳琦1,舒林径2,熊毅3,张艺馨1,向琳3,伍颖颖3()   

  1. 1.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心,四川 成都(610041)
    2.重庆医科大学附属口腔医院种植科,重庆(400016)
    3.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院种植科,四川 成都(610041)
  • 收稿日期:2019-05-15 修回日期:2019-09-16 出版日期:2020-01-20 发布日期:2020-01-17
  • 通讯作者: 伍颖颖
  • 作者简介:周佳琦,学士,Email: zhoujiaqi1013@163.com
  • 基金资助:
    四川省科技计划项目(2018RZ0088);四川省科技计划项目(2018RZ0087);国家自然科学基金项目(81701007)

Study on the role of FoxO1 in the regulation of osteoblastic metabolism by 1,25(OH)2D3 in a high glucose environment

ZHOU Jiaqi1,SHU Linjing2,XIONG Yi3,ZHANG Yixin1,XIANG Lin3,WU Yingying3()   

  1. 1.State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
    2.Department of Oral Implantology, Stomatological Hospital of Chongqing Medical University, Chongqing 400016, China
    3.Department of Oral Implantology, West China Hospital of Stomatology, State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, Sichuan University, Chengdu 610041, China
  • Received:2019-05-15 Revised:2019-09-16 Online:2020-01-20 Published:2020-01-17
  • Contact: Yingying WU

摘要:

目的 探究1,25(OH)2D3对高糖环境下骨代谢的调控作用,为1,25(OH)2D3对高糖环境下成骨细胞的可能调控机制提供证据。方法 将成骨细胞系MC3T3-E1细胞分3组培养:①对照组,低糖(5.5 mmol/L)DMEM培养基培养;②高糖组:高糖(22 mmol/L)DMEM培养基培养;③高糖+1,25(OH)2D3组:高糖DMEM+1,25(OH)2D3 培养基培养。利用CCK-8法检测各组细胞增殖活性;Annexin V, FITC凋亡试剂盒检测细胞凋亡情况;茜素红半定量分析细胞分化水平;分别通过qRT-PCR检测叉头转录因子-1(forkhead transcription factor 1,FoxO1)mRNA表达,免疫荧光观察FoxO1蛋白表达变化和其与细胞核的相对位置关系。结果 相较于对照组,高糖组成骨细胞过度增殖、凋亡显著增加、成骨分化受抑制(P < 0.05),FoxO1 mRNA增加(P=0.006),FoxO1蛋白核内转移增加(P=0.001)。相较于高塘组,高糖+1,25(OH)2D3 组细胞的过度增殖受到抑制、凋亡减少、成骨分化较高糖组改善(P < 0.05),FoxO1 mRNA减少(P=0.006),FoxO1蛋白核内转移受阻(P < 0.001)。结论 1,25(OH)2D3可能通过降低高糖环境中成骨细胞FoxO1 mRNA表达,阻止FoxO1向细胞核内转移,抑制高糖环境下成骨细胞的异常增殖、凋亡,并逆转高糖对成骨分化的抑制作用。

关键词: 成骨细胞, 成骨分化, 高糖血症, 糖尿病, 叉头转录因子-1, 增殖, 凋亡, 1,25(OH)2D3, 维生素D, 骨代谢

Abstract:

Objective To explore the effect of 1,25(OH)2D3 on the regulation of bone metabolism in a high-glucose environment and to provide evidence for the possible regulatory mechanism of 1,25(OH)2D3 on osteoblasts in a high-glucose environment.Methods The osteoblast cell line MC3T3-E1 was cultured in 3 groups: ① control group, cultured in low-glucose (5.5 mmol/L) DMEM; ② high-glucose group: cultured in high-glucose (22 mmol/L) DMEM; ③ high-glucose +1,25(OH)2D3 group: high-glucose DMEM + 1,25(OH)2D3 medium culture. The CCK-8 method was used to detect cell proliferation in each group; Annexin V and FITC apoptosis kits were used to detect apoptosis; Alizarin red was used to semiquantitatively analyze cell differentiation; qRT-PCR was used to detect forkhead transcription factor-1 (forkhead transcription factor 1, FoxO1) mRNA expression. Immunofluorescence was used to observe the changes in FoxO1 protein expression and its relative position in the nucleus.Results Our analysis showed that compared with those in the control group, the osteoblast apoptosis and proliferation in the high-glucose group were improved, while differentiation was inhibited (P < 0.05); at the same time, the mRNA expression of FoxO1(P = 0.006) was reduced. The immunofluorescence results showed that more FoxO1 was inside the nucleus (P < 0.001). Compared with those in the high-glucose group, excessive proliferation was inhibited, apoptosis was reduced, and osteogenic differentiation was improved in the high-glucose +1,25(OH)2D3 group (P < 0.05); furthermore, FoxO1 mRNA was decreased (P = 0.006), and the transfer of FoxO1 protein was blocked (P < 0.001).Conclusion We found that 1,25(OH)2D3 may prevent the transfer of FoxO1 to the cell nucleus, inhibit the abnormal proliferation and apoptosis of osteoblasts in a high-glucose environment, and reverse the inhibitory effect of high glucose on the differentiation of osteoblasts.

Key words: osteoblast, osteogenic differentiation, hyperglycemia, diabetes, forkhead transcription factor 1(FoxO1), proliferation, apoptosis, 1,25(OH)2D3, vitamin D, bone metabolism

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