糖尿病大鼠牙髓干细胞增殖及成牙/成骨分化能力研究

›› 2018, Vol. 9 ›› Issue (1): 20-23.

糖尿病大鼠牙髓干细胞增殖及成牙/成骨分化能力研究

王燕萍¹,陆乐¹,徐云龙²,于金华²

1. 苏州大学附属口腔医院，苏州口腔医院
2. 南京医科大学口腔医学院

收稿日期:2017-11-20 修回日期:2018-01-15 出版日期:2018-03-25 发布日期:2018-04-04
通讯作者: 王燕萍 E-mail:wangyp0512@163.com
基金资助:
江苏省口腔疾病重点实验室开放课题

The proliferation and odonto/osteogenic differentiation of dental pulp stem cells from diabetic rats

Received:2017-11-20 Revised:2018-01-15 Online:2018-03-25 Published:2018-04-04
Supported by:
The fund of the key laboratory of jiangsu province

摘要/Abstract

摘要： 目的：探讨高血糖大鼠牙髓干细胞（DPSCs）增殖及成牙/成骨分化能力。方法：选取3个月龄糖尿病模型动物GK大鼠及正常对照Wistar大鼠，测定尾静脉血糖值，待血糖稳定1周后，通过酶消化法分离培养两组大鼠DPSCs，通过MTT法测定两组大鼠DPSCs生长曲线，实时定量RT-PCR及Western blot检测两组大鼠DPSCs牙本质基质蛋白1（DMP1）、Osterix（Osx）、骨钙素（OCN）等成牙/成骨向分化指标的表达。结果：GK大鼠餐后2小时尾静脉血糖值稳定在20.1~22.3 mmol/L，Wistar大鼠为5.8~6.3 mmol/L。MTT结果显示，GK大鼠DPSCs增殖能力从培养第3天到第7天均较Wistar大鼠DPSCs低（P<0.05）。实时定量RT-PCR及Western blot检测结果显示GK大鼠DPSCs成牙/成骨向分化指标的表达明显较正常对照Wistar大鼠低。结论：高血糖大鼠DPSCs增殖能力及成牙/成骨向分化能力均较正常大鼠低。

Abstract: Objective：To explore the proliferation and odonto/osteogenic potential of DPSCs separated from GK rat with hyperglycemia. Methods： The 3-month-old GK rat (one diabetic model rat) and Wistar rat were selected. The DPSCs were isolated and cultured when the blood glucose level had been stable for 1 weeks. MTT assay was applied to estimate the proliferation of the two DPSCs. The odonto/osteogenic markers (OCN, OSX and DMP1) of the two DPSCs were measured by real-time RT-PCR and Western blot. Results： The level of blood glucose in GK rats remained about 20.1~22.3 mmol/L (high glucose) and the level of blood glucose of Wistar rats was 5.8 ~ 6.3 mmol/L (normal glucose level). MTT assay showed the proliferative activity of DPSCs from GK was significantly lower than that of the Wistar rat. Moreover, the expressions of OCN, OSX and DMP1 in DPSCs of GK rats were significantly weaker than those in Wistar rats. Conclusions：The DPSCs from the donor with hyperglycemia or diabetes exhibited the weaker proliferation and odonto/osteogenic potential.

王燕萍陆乐徐云龙于金华. 糖尿病大鼠牙髓干细胞增殖及成牙/成骨分化能力研究[J]. , 2018, 9(1): 20-23.

参考文献

[1]Gronthos S, Mankani M, Brahim J, et al.Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo[J].Proc Natl Acad Sci U S A, 2000, 97(25):13625-13630
[2]Wang Y, Zheng Y, Wang Z, et al.10(-7) m 17β-oestradiol enhances odonto/osteogenic potency of human dental pulp stem cells by activation of the NF-κB pathway[J].Cell Prolif, 2013, 46(6):677-684
[3]Wang Y, Yan M, Yu Y, et al.Estrogen deficiency inhibits the odonto/osteogenic differentiation of dental pulp stem cells via activation of the NF-κB pathway[J].Cell Tissue Res, 2013, 352(3):551-559
[4]Wang Y, Yan M, Fan Z, et al.Mineral trioxide aggregate enhances the odonto/osteogenic capacity of stem cells from inflammatory dental pulps via NF-κB pathway[J].Oral Dis, 2014, 20(7):650-658
[5]Al-Habib M, Yu Z, Huang GT.Small molecules affect human dental pulp stem cell properties via multiple signaling pathways[J].Stem Cells Dev, 2013, 22(17):2402-2413
[6]Cao B, Liu N, Wang W.High glucose prevents osteogenic differentiation of mesenchymal stem cells via lncRNA AK028326/CXCL13 pathway[J].Biomed Pharmacother, 2016, 84:544-551
[7]Kato H, Taguchi Y, Tominaga K, et al.High Glucose Concentrations Suppress the Proliferation of Human Periodontal Ligament Stem Cells and Their Differentiation Into Osteoblasts[J].J Periodontol, 2016, 87(4):e44-e51
[8]Lamster IB, Pagan M.Periodontal disease and the metabolic syndrome[J].Int Dent J, 2017, 67(2):67-77
[9]McClelland Descalzo DL, Satoorian TS, Walker LM, et al.Glucose-Induced Oxidative Stress Reduces Proliferation in Embryonic Stem Cells via FOXO3A/β-Catenin-Dependent Transcription of p21(cip1)[J].Stem Cell Reports, 2016, 7(1):55-68
[10]Zhang B, Liu N, Shi H, et al.High glucose microenvironments inhibit the proliferation and migration of bone mesenchymal stem cells by activating GSK3β[J].J Bone Miner Metab, 2016, 34(2):140-150
[11]Lee MH, Kwon TG, Park HS, et al.BMP-2-induced Osterix expression is mediated by Dlx5 but is independent of Runx2[J].Biochem Biophys Res Commun, 2003, 309(3):689-694
[12]段瑞平, 吴凌, 林云锋, 等.不同成骨诱导作用下骨髓间充质干细胞的基因表达模式[J].四川大学学报(医学版), 2006, 37(6):856-859
[13]Wu G, Deng ZH, Fan XJ, et al.Odontogenic potential of mesenchymal cells from hair follicle dermal papilla[J].Stem Cells Dev, 2009, 18(4):583-589
[14]Nakashima M, Tachibana K, Iohara K, et al.Induction of reparative dentin formation by ultrasound-mediated gene delivery of growth/differentiation factor 11[J].Hum Gene Ther, 2003, 14(6):591-597