ephrinB2基因转染脱落乳牙来源的牙髓干细胞提高其成骨/成牙本质分化的研究  

Transgenic expression of ephrinB2 in stem cells from human exfoliated deciduous teeth enhances osteogenic differentiation

作　　者：胡晓燕[1] 朱绍跃 徐建光[1] 李午丽[1] 张菁[1] Hu Xiaoyan;Zhu Shaoyue;Xu Jianguang(Stomatology College of Anhui Medical University,The Affiliated Stomatological Hospital of Anhui Medical University,Key Lab.of Oral Diseases Research of Anhui Province,Hefei 230032;Affiliated Xuzhou Stomatological Hospital of Xuzhou Medical University,Xuzhou 221002)

机构地区：[1]安徽医科大学口腔医学院安徽医科大学附属口腔医院安徽省口腔疾病研究重点实验室,合肥230032 [2]徐州医科大学附属徐州口腔医院,徐州221002

出　　处：《安徽医科大学学报》2019年第4期564-569,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽高校自然科学研究项目(编号:KJ2016A324;KJ2018A0199);安徽省自然科学基金(编号:1508085SQH222)

摘　　要：目的观察促红细胞生成素肝细胞激酶受体B4/促红细胞生成素肝细胞激酶受体配体B2 (EphB4/ephrinB2)信号通路在调控过表达ephrinB2的脱落乳牙来源牙髓干细胞(SHEDs)成骨/成牙本质分化中的作用。方法从滞留乳牙中分离出SHEDs,用hEfnB2-GFP-Bsd载体和空载体(GFP-Bsd)转染后进行成骨/成牙本质分化诱导。用反转录聚合酶链反应(RT-PCR)、碱性磷酸酶实验(ALP)实验、茜素红S染色实验来检测EfnB2-SHEDs组、空载体对照组(Vector-SHEDs)的成骨分化效果,并用Western blot、免疫沉淀和免疫共沉淀试验来检测ephrinB2和EphB4磷酸化蛋白表达水平。结果 ALP实验和茜素红S染色实验证实:EfnB2-SHEDs组比Vector-SHEDs组表现出更高的ALP活性和矿化能力。RT-PCR显示:EfnB2-SHEDs组成牙本质/成骨标志物的表达明显高于Vector-SHEDs组,在成骨分化中,EphB4受体通过磷酸化被活化。结论基因转染ephrinB2能够通过刺激ephrinB2和EphB4的磷酸化来提高SHEDs的成骨/成牙本质分化。Objective To investigate the role of ephrinB2/EphB4 signaling in the osteogenic/odontogenic differentiation of SHEDs via over-expression ephrinB2 in SHEDs.Methods SHEDs were isolated from exfoliated deciduous teeth and transfected with hEfnB2-GFP-Bsd(GFP-Bsd as empty Vector).The transfected SHEDs were then induced into osteo-/odontoblasts under special differentiation medium.RT-PCR,Alizarin-red S staining,and ALP assay were used to analyze the effect of osteogenic/odontogenic differentiation of EfnB2-SHEDs and Vector-SHEDs.The phosphorylated protein expression level of ephrinB2 and EphB4 were invested via Western blot,immunoprecipitation assays and co-immunoprecipitation assays.Results Compared with Vector-SHEDs,EfnB2-SHEDs possessed higher ALP activity and mineralization capacity.The odonto-/osteogenic genes expression were significantly enhanced in EfnB2-SHEDs group,and the activity of EphB4 receptor was triggered through phosphorylation in odonto-/osteogenic differentiation.Conclusion Our study shows that the odonto-/osteogenic differentiation of SHEDs can be improved through stimulation of the phosphorylation of ephrinB2/Ephb4.

关 键 词：乳牙牙髓干细胞 EPHRINB2 基因转染 成骨分化 

分 类 号：R781.3[医药卫生—口腔医学]