IGF-1促进大鼠上皮根鞘HERS细胞成牙骨质分化的研究

›› 2020, Vol. 11 ›› Issue (1): 9-12.

IGF-1促进大鼠上皮根鞘HERS细胞成牙骨质分化的研究

王世恺¹,李雪冰¹,陈国庆¹,田卫东²

1. 四川大学华西口腔医院
2. 四川大学华西口腔医学院

收稿日期:2019-11-04 修回日期:2020-02-24 出版日期:2020-03-25 发布日期:2020-04-07
通讯作者: 田卫东 E-mail:drtwd@sina.com

IGF-1 enhance the cementoblastic differentiation of Hertwig’s epithelial root sheath cells of rats

Received:2019-11-04 Revised:2020-02-24 Online:2020-03-25 Published:2020-04-07
Contact: Wei-Dong TIAN E-mail:drtwd@sina.com

摘要/Abstract

摘要： 目的：研究胰岛素样生长因子1（IGF-1）对赫特维希上皮根鞘（HERS）细胞增殖和分化的影响。方法：取出生后8 d的SD大鼠，分离纯化HERS上皮细胞，分为对照组和IGF-1干预组；CCK8检测细胞生长曲线；免疫荧光染色检测上皮细胞标记物细胞角蛋白14（CK14）、上皮细胞钙黏蛋白（E-cadherin）和间充质细胞标记物波形蛋白（Vimentin）的表达；实时定量RT-PCR检测上皮细胞标记物E-cadherin，间充质细胞标记物Vimentin、神经钙黏蛋白（N-cadherin）、Ⅰ型胶原蛋白（Col1），成牙骨质分化相关基因骨涎蛋白（Bsp）、Runt相关转录因子2（Runx2）、碱性磷酸酶（Alp）等基因表达水平；茜素红染色检测细胞矿化情况。结果：HERS细胞能够同时表达上皮及间充质细胞标记物；IGF-1干预后，免疫荧光染色及实时定量RT-PCR结果显示HERS细胞上皮细胞标记物表达显著下降，间充质细胞标记物表达显著上调，并且明显促进细胞增殖（P＜0.05）。矿化诱导环境下，IGF-1干预组7、14 d均显示成牙骨质分化相关基因表达显著上调，促进矿化结节形成（P＜0.05）。结论：IGF-1诱导促进HERS细胞增殖、上皮-间充质转化和成牙骨质细胞向分化

关键词: 赫特维希上皮根鞘, 胰岛素样生长因子1, 成牙骨质细胞

Abstract: Objective:To study the effects of IGF-1 on the proliferation and differentiation of Hertwig’s epithelial root sheath(HERS) in vitro.Methods:HERS cells were isolated and cultured in vitro, and were treated with IGF-1. Proliferation of HERS cells was evaluated by CCK8 assay. Immunofluorescence staining detected the expression of CK14, E-cadherin, Vimentin of HERS cells. Real-time PCR detected the expression of E-cadherin, N-cadherin, Vimentin, Bsp,Col1, Runx2, Alp. And osteogenic differentiation of HERS cells was determined by Alizarin red staining.Results:HERS cells could have the expression of the marker gene of epithelia and mesenchyme.IGF-1 treatment significantly decreased the expression of epithelial marker gene and increased the expression of mesenchymal marker gene and enhanced the cell proliferation. HERS cells culturedin mineralization-induction medium showed significantly upregulated the expression of genes related to the cementoblastic differentiation such as Bsp,Col1, Runx2, Alp on day 7 and day 14(P＜0.05). IGF-1 treatment also significantly enhanced the mineralization nodule formation of HERS cells. Conclusions:IGF-1 regulated the proliferation, cementoblastic differentiation and EMT of HERS cells.

Key words: Hertwig’s epithelial root sheath, IGF-1, Cementoblast

王世恺李雪冰陈国庆田卫东. IGF-1促进大鼠上皮根鞘HERS细胞成牙骨质分化的研究[J]. 口腔生物医学, 2020, 11(1): 9-12.

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