FoxO1在1,25（OH）2D3调控高糖环境下成骨细胞代谢中的作用  

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

作　　者：周佳琦 舒林径 熊毅 张艺馨 向琳[3] 伍颖颖[3] ZHOU Jiaqi;SHU Linjing;XIONG Yi;ZHANG Yixin;XIANG Lin;WU Yingying(State Key Lab⁃oratory of Oral Diseases,National Clinical Research Center for Oral Diseases,West China Hospital of Stomatology,Sich⁃uan University,Chengdu 610041,China;Department of Oral Implantology,Stomatological Hospital of Chongqing Medical University,Chongqing 400016,China;Department of Oral Implantology,West China Hospital of Stomatolo⁃gy,State Key Laboratory of Oral Diseases,National Clinical Research Center for Oral Diseases,Sichuan University,Chengdu 610041,China)

机构地区：[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

出　　处：《口腔疾病防治》2020年第1期24-29,共6页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：四川省科技计划项目(2018RZ0088,2018RZ0087);国家自然科学基金项目(81701007)

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。ObjectiveTo explore the effect of 1,25(OH)2D3 on the regulation of bone metabolism in a high-glucoseenvironment and to provide evidence for the possible regulatory mechanism of 1,25(OH)2D3 on osteoblasts in a high-glu-cose environment.MethodsThe osteoblast cell line MC3 T3-E1 was cultured in 3 groups:(1) control group,culturedin low-glucose(5.5 mmol/L) DMEM;(2) high-glucose group:cultured in high-glucose(22 mmol/L) DMEM;(3) high-glu-cose +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 usedto semiquantitatively analyze cell differentiation;q RT-PCR was used to detect forkhead transcription factor-1(forkheadtranscription factor 1,FoxO1) m RNA expression.Immunofluorescence was used to observe the changes in FoxO1 pro-tein expression and its relative position in the nucleus.ResultsOur analysis showed that compared with those in thecontrol group,the osteoblast apoptosis and proliferation in the high-glucose group were improved,while differentiationwas inhibited(P < 0.05);at the same time,the m RNA expression of FoxO1(P = 0.006) was reduced.The immunofluores-cence results showed that more FoxO1 was inside the nucleus(P < 0.001).Compared with those in the high-glucosegroup,excessive proliferation was inhibited,apoptosis was reduced,and osteogenic differentiation was improved in thehigh-glucose +1,25(OH)2D3 group(P < 0.05);furthermore,FoxO1 m RNA was decreased(P = 0.006),and the transfer ofFoxO1 protein was blocked(P < 0.001).ConclusionWe found that 1,25(OH)2D3 may prevent the transfer of FoxO1 tothe cell nucleus,inhibit the abnormal proliferation and apoptosis of osteoblasts in a high-glucose environment,and re-verse the inhibitory effect of high glucose on the differentiation of osteoblasts.

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 

分 类 号：R78[医药卫生—口腔医学]