GLUT1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响

doi:10.13701/j.cnki.kqyxyj.2024.10.014

口腔医学研究 ›› 2024, Vol. 40 ›› Issue (10): 920-927.DOI: 10.13701/j.cnki.kqyxyj.2024.10.014

GLUT1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响

刘贞^*, 许针针, 相黎黎, 崔颖颖, 孙轶群

滨州医学院附属医院儿童口腔科山东滨州 256600

收稿日期:2024-07-10 出版日期:2024-10-28 发布日期:2024-10-24
通讯作者: *刘贞,E-mail:18254386775@163.com
作者简介:刘贞(1986~),女,山东济宁人,硕士,主治医师,研究方向:儿童牙齿生长发育,儿童口腔疾病的预防及治疗。

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

LIU Zhen^*, XU Zhenzhen, XIANG Lili, CUI Yingying, SUN Yiqun

Department of Pediatric Dentistry, Binzhou Medical University Hospital, Binzhou 256600, China

Received:2024-07-10 Online:2024-10-28 Published:2024-10-24

摘要/Abstract

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。

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

Abstract: 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 an important role in early tooth development in mice.

Key words: glucose transporter 1, glucose transporter 2, glucose uptake, early development of teeth, phloretin

刘贞, 许针针, 相黎黎, 崔颖颖, 孙轶群. GLUT1、GLUT2介导的葡萄糖摄取对小鼠牙齿早期发育的影响[J]. 口腔医学研究, 2024, 40(10): 920-927.

LIU Zhen, XU Zhenzhen, XIANG Lili, CUI Yingying, SUN Yiqun. Effects of GLUT1 and GLUT2 Mediated Glucose Uptake on Early Tooth Development in Mice[J]. Journal of Oral Science Research, 2024, 40(10): 920-927.

参考文献

[1] Brăescu R, Săvinescu SD, Tatarciuc MS, et al. Pointing on the early stages of maxillary bone and tooth development-histological findings[J]. Rom J Morphol Embryol, 2020, 61(1):167-174.
[2] Truong TTK, Fujii S, Nagano R, et al. Arl4c is involved in tooth germ development through osteoblastic/ameloblastic differentiation[J]. Biochem Biophys Res Commun, 2023, 679:167-174.
[3] Park Y, Lee HJ, Sim DY, et al. Inhibition of glycolysis and SIRT1/GLUT1 signaling ameliorates the apoptotic effect of Leptosidin in prostate cancer cells[J]. Phytother Res, 2024, 38(3):1235-1244.
[4] Sun B, Chen H, Xue JS, et al. The role of GLUT2 in glucose metabolism in multiple organs and tissues[J]. Mol Biol Rep,2023,50(8):6963-6974.
[5] Ohshima H, Wartiovaara J, Thesleff I, et al. Developmental regulation and ultrastructure of glycogen deposits during murine tooth morphogenesis[J]. Cell Tissue Res,1999, 297(2):271-281.
[6] 陈书盈,孙娟,王茂颖.SLC2A1,SLC4A4以及ADCY5基因在大鼠牙胚发育中的表达研究[J].临床口腔医学杂志,2023,39(7):387-391.
[7] Ida-Yonemochi H, Nakatomi M, Harada H, et al. Glucose uptake mediated by glucose transporter 1 is essential for early tooth morphogenesis and size determination of murine molars[J]. Dev Biol, 2012, 363(1):52-61.
[8] Grobholz R, Hacker HJ, Thorens B, et al. Reduction in the expression of glucose transporter protein GLUT 2 in preneoplastic and neoplastic hepatic lesions and reexpression of GLUT1 in late stages of hepatocarcinogenesis[J]. Cancer Res, 1993, 53(18):4204-4211.
[9] Baba O. Production of monoclonal antibody that recognizes glycogen and its application for immunohistochemistry[J]. Kokubyo Gakkai Zasshi, 1993, 60(2):264-287.
[10] Liu M, Yang Q, Zuo HY, et al. Dynamic patterns of histone lactylation during early tooth development in mice[J]. J Mol Histol, 2023, 54(6):665-673.
[11] Littleflower AB, Antony GR, Parambil ST, et al. Metabolic phenotype intricacies on altered glucose metabolism of breast cancer cells upon glut-1 inhibition and mimic hypoxia in vitro[J]. Appl Biochem Biotechnol, 2023, 195(10):5838-5854.
[12] Sibiak R, Ozegowska K, Wender-Ozegowska E, et al. Fetomaternal expression of glucose transporters (GLUTs)-biochemical, cellular and clinical aspects[J]. Nutrients, 2022, 14(10):2025.
[13] Ahn YS, Zerban H, Grobholz R, et al. Sequential changes in glycogen content, expression of glucose transporters and enzymic patterns during development of clear/acidophilic cell tumors in rat kidney[J]. Carcinogenesis, 1992, 13(12):2329-2334.
[14] Kuroki S, Yokoo S, Terashi H, et al. Epithelialization in oral mucous wound healing in terms of energy metabolism[J]. Kobe J Med Sci, 2009, 55(1):E5-E15.
[15] Grobholz R, Hacker HJ, Thorens B, et al. Reduction in the expression of glucose transporter protein GLUT 2 in preneoplastic and neoplastic hepatic lesions and reexpression of GLUT 1 in late stages of hepatocarcinogenesis[J]. Cancer Res, 1993, 53(18):4204-4211.
[16] Songyang YY, Li W, Li WQ, et al. The inhibition of GLUT1-induced glycolysis in macrophage by phloretin participates in the protection during acute lung injury[J]. Int Immunopharmacol, 2022, 110:109049.
[17] Kim JY, Cha YG, Cho SW, et al. Inhibition of apoptosis in early tooth development alters tooth shape and size[J]. J Dent Res, 2006, 85(6):530-535.
[18] Ida-Yonemochi H, Otsu K, Harada H, et al. Functional expression of sodium-dependent glucose transporter in amelogenesis[J]. J Dent Res, 2020, 99(8):977-986.

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

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