IGF-1促进大鼠上皮根鞘HERS细胞成牙骨质分化的研究  

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

作　　者：王世恺 李雪冰 陈国庆 田卫东[1] WANG Shikai;LI Xuebing;CHEN Guoqing;TIAN Weidong(Department of Oral and Maxillofacial Surgery,National Engineering Laboratory for Oral Regenerative Medicine,West China Hospital of Stomatology,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西口腔医院创伤整形外科,四川大学华西口腔医院口腔再生医学国家地方联合工程实验室,四川成都610041

出　　处：《口腔生物医学》2020年第1期9-12,共4页Oral Biomedicine

基　　金：国家自然科学基金(31600789)。

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RTPCR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。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 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.

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 

分 类 号：R780.2[医药卫生—口腔医学]