干旱区生态环境知识资源中心(Arid): 高糖损害小鼠胚胎成骨前体细胞糖代谢并通过原纤毛/Hedgehog信号通路抑制成骨分化的研究

Arid

	高糖损害小鼠胚胎成骨前体细胞糖代谢并通过原纤毛/Hedgehog信号通路抑制成骨分化的研究
其他题名	High glucose impairs glucose metabolism and inhibits osteoblast differentiation via primary cilia/Hedgehog pathway in mouse embryonic osteoblast precursor cells
	刘杰; 许洁; 陈茜; 汪长东
来源期刊	中国糖尿病杂志
ISSN	1006-6187
出版年	2021
卷号	29 期号:11 页码:836-842
中文摘要	目的研究高糖对小鼠胚胎成骨前体细胞(MC3T3-E1)增殖、成骨分化和糖代谢影响。方法MC3T3-E1细胞予5.6 mmol/L(Con组)、25.0 mmol/L(HG组)葡萄糖浓度的alpha-MEM培养。 CCK-8、Ki67免疫荧光检测细胞增殖,免疫荧光检测原纤毛;ELISA检测碱性磷酸酶(ALP)活力;Western blot检测ALP、骨桥蛋白(OPN)、乙酰化alpha-tubulin(Ac-alpha-tubulin)、gamma-tubulin、印度/音/沙漠刺猬信号分子(IHH,SHH,DHH)、神经胶质瘤相关癌基因(Gli1/Gli2)、糖原合酶1(GYS1)、GluT4、磷酸果糖激酶(PFKM)、乙酰-CoA羧化酶alpha(ACACA)、糖原磷酸化酶(PYGL)、丙酮酸脱氢酶磷酸酶1(PDP1)蛋白表达;茜素红染色检测矿化结节。结果与Con组比较,HG组Ki67阳性率升高[(13.852.28)% vs(30.233.83)%,P<0.01],矿化结节形成受抑制(P<0.01),ALP酶活力降低(P<0.01),ALP、OPN、Gli2、IHH、SHH、GYS1、ACACA、PFKM、GluT4、Ac-alpha-tubulin、gamma-tubulin蛋白表达减低(P<0.05或P<0.01),PYGL、PDP1表达升高(P<0.05或P<0.01),原纤毛Ac-alpha-tubulin和gamma-tubulin平均荧光强度降低(P<0.05或P<0.01)。结论高糖促进MC3T3-E1细胞增殖,损害糖代谢机制,破坏原纤毛,阻断Hedgehog信号,抑制成骨分化和矿化。
英文摘要	Objective To study the effects of high glucose on the proliferation,osteoblast differentiation and glucose metabolism of mouse embryonic osteoblast precursor cells(MC3T3-E1).Methods MC3T3-E1 cells were cultured with glucose alpha-MEM at concentrations of 5.6 mmol/L(Con group)and 25.0 mmol/L(HG group).Cell proliferation was detected by CCK-8 and Ki67 immunofluorescence.Primary cilia were detected by immunofluorescence.ELISA assayed alkaline phosphatase(ALP)activity.Western blot determined protein expression of ALP,osteopontin(OPN),gamma-tubulin,acetylated alpha-tubulin(Ac-alpha-tubulin),Indian/Sonic/Desert hedgehog signaling molecule(IHH,SHH,DHH),glioma-associated oncogene(Gli1,Gli2),glycogen synthase 1(GYS1),glucose transporter 4(GluT4),phosphofructokinase(PFKM),acetyl-CoA carboxylase alpha(ACACA),glycogen phosphorylase(PYGL)and pyruvate dehydrogenase phosphatase 1(PDP1).Alizarin red staining detected extracellular matrix mineralized nodules.Results Compared with Con group,the positive rate of Ki67 in HG group increased[(13.852.28)% vs(30.233.83)%,P<0.01],the formation of mineralized nodules was inhibited(P<0.01),and ALP activity decreased(P<0.01).The expression of ALP,OPN,Gli2,IHH,SHH,GYS1,ACACA,PFKM,GluT4,Ac-alpha-tubulin,gamma-tubulin decreased(P<0.05 or P<0.01),the expression of PYGL and PDP1 increased(P<0.05 or P<0.01),and the average fluorescence intensity of Ac-alpha-tubulin and gamma-tubulin of primary cilia decreased(P<0.05 or P<0.01).Conclusion High glucose promotes the proliferation of MC3T3-E1 cells,but disorders glucose metabolism,impairs primary cilia,blocks Hedgehog signaland inhibits osteoblast differentiation and mineralization.
中文关键词	高糖 ; 成骨分化 ; 原纤毛 ; 糖代谢
英文关键词	High glucose Osteoblast differentiation Primary cilia Glucose metabolism
类型	Article
语种	中文
收录类别	CSCD
WOS类目	General & Internal Medicine
CSCD记录号	CSCD:7088500
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/378140
作者单位	刘杰, 重庆医科大学基础医学院, 400016.; 许洁, 重庆医科大学基础医学院, 400016.; 陈茜, 重庆医科大学基础医学院, 400016.; 汪长东, 重庆医科大学基础医学院, 400016.
推荐引用方式 GB/T 7714	刘杰,许洁,陈茜,等. 高糖损害小鼠胚胎成骨前体细胞糖代谢并通过原纤毛/Hedgehog信号通路抑制成骨分化的研究[J],2021,29(11):836-842.
APA	刘杰,许洁,陈茜,&汪长东.(2021).高糖损害小鼠胚胎成骨前体细胞糖代谢并通过原纤毛/Hedgehog信号通路抑制成骨分化的研究.中国糖尿病杂志,29(11),836-842.
MLA	刘杰,et al."高糖损害小鼠胚胎成骨前体细胞糖代谢并通过原纤毛/Hedgehog信号通路抑制成骨分化的研究".中国糖尿病杂志 29.11(2021):836-842.