GLUT1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响  

Effects of GLUT1 and GLUT2 Mediated Glucose Uptake on Early Tooth Development in Mice

作　　者：刘贞 许针针[1] 相黎黎 崔颖颖 孙轶群[1] LIU Zhen;XU Zhenzhen;XIANG Lili;CUI Yingying;SUN Yiqun(Department of Pediatric Dentistry,Binzhou Medical University Hospital,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院儿童口腔科,山东滨州256600

出　　处：《口腔医学研究》2024年第10期920-927,共8页Journal of Oral Science Research

摘　　要：目的:探究葡萄糖转运蛋白(glucose transporter,GLUT)1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响。方法:收集胚胎13.5 d(embryonic day 0.5,E13.5)、E14.5、E16.5、E18.5和出生后1 d(postnatal day 1,P1)时期的下颌磨牙牙胚、上颌切牙牙胚;实时荧光定量聚合酶链反应(real time-quantitative polymerase chain reaction,RT-qPCR)和Western blot检测牙胚中Glut1、Glut2 mRNA和蛋白水平;免疫组织化学染色检测牙胚中GLUT1、GLUT2、Ki67、糖原水平。将下颌磨牙牙胚在无糖DMEM培养基、高糖且含不同浓度的根皮素(0、0.25、0.5 mmol/L)的DMEM培养基中培养9 d;并对其进行苏木精-伊红(hematoxylin-eosin,HE)染色。结果:(1)在E13.5时期,GLUT1在成釉器中高表达,细胞增殖活跃,糖原在牙板和牙囊中大量沉积;在E14.5、E18.5时期,GLUT1在成釉器中表达逐渐降低,细胞增殖减少,糖原在成釉器和牙乳头中大量沉积;在P1时期,GLUT1在中间层和内釉上皮中表达较多,细胞增殖增多,糖原在牙乳头中少量沉积。在整个牙胚发育阶段,GLUT2表达相对较少。(2)GLUT1在前成釉细胞、前成牙本质细胞中高表达,而在分化后的成釉细胞、成牙本质细胞中表达较少;GLUT2与GLUT1呈现相反的表达趋势。(3)0.5 mmol/L根皮素能够抑制E13.5时期外植体牙胚发育,0.25 mmol/L根皮素不抑制E13.5、E14.5时期外植体牙胚发育,但能够导致牙胚变小,且具有根皮素浓度依赖性。无糖培养基能够抑制E13.5、E14.5时期外植体牙胚发育。结论:GLUT1、GLUT2在牙齿早期发育中的表达受到精确的时空调控,由GLUT1、GLUT2介导的葡萄糖摄取在小鼠牙齿早期发育中发挥重要作用。Objective:To explore the effects of glucose transporter(GLUT)1 and GLUT2 mediated glucose uptake on early tooth development in mice.Methods:The mandibular molar tooth germ and maxillary incisor tooth germ during embryonic day 13.5(E13.5),E14.5,E16.5,E18.5,and postnatal day 1(P1)were collected.RT-qPCR and Western blot were used to detect Glut1 and Glut2 mRNA and protein levels in tooth germ.Immunohistochemical staining was used to detect GLUT1,GLUT2,Ki67,and glycogen levels in tooth germ.The mandibular molar tooth germ was cultivated in glucose-free DMEM medium and high glucose DMEM medium with different concentrations of Phloretin(0 mmol/L,0.25 mmol/L,0.5 mmol/L)for 9 days.Results:(1)During E13.5,GLUT1 was strongly expressed in enamel organ,with active cell proliferation and abundant deposition of glycogen in dental lamina and odontotheca.During E14.5 and E18.5,the expression of GLUT1 was gradually decreased in enamel organ,cell proliferation was decreased,and glycogen was heavily deposited in enamel organ and dental papillae.During P1,GLUT1 was highly expressed in middle layer and inner enamel epithelium,with increased cell proliferation and minimal deposition of glycogen in dental papilla.During the entire stage of tooth germ development,GLUT2 expression was relatively low.(2)GLUT1 was strongly expressed in preameloblasts and preodontoblasts cells,but less expressed in differentiated ameloblasts and odontoblasts cells.GLUT2 and GLUT1 exhibited opposite expression trends.(3)0.5 mmol/L Phloretin could inhibit the development of tooth germ in E13.5 stage,while 0.25 mmol/L root bark extract didn’t inhibit the development of tooth germ in E13.5 and E14.5 stage,but could cause tooth germ to shrink and had a concentration dependent effect on Phloretin.Glucose-free culture medium could inhibit the development of explant dental germ in E13.5 and E14.5 stages.Conclusion:The expression of GLUT1 and GLUT2 in early tooth development were precisely spatiotemporal regulated,and GLUT1 and GLUT2 mediated glucose uptake played

关 键 词：葡萄糖转运蛋白1 葡萄糖转运蛋白2 葡萄糖摄取 牙齿早期发育 根皮素 

分 类 号：R781[医药卫生—口腔医学]