Notch信号通路和miR-21在周围神经损伤修复中的交互作用机制  

Interactions of Notch signaling pathway and miR-21 in peripheral nerve repair

作　　者：杨凯云 孙佩佩 武文亮[1] 刘海春[1] Yang Kai-yun;Sun Pei-pei;Wu Wen-liang;Liu Hai-chun(Department of Orthopedics,Shandong University Qilu Hospital,Jinan 250012,Shandong Province,China)

机构地区：[1]山东大学齐鲁医院骨外科,山东省济南市250012

出　　处：《中国组织工程研究》2018年第32期5139-5144,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省重点研发计划(2017GSF218047)~~

摘　　要：背景:神经损伤修复是外科临床和基础研究的主要热点之一。小间隙套管治疗周围神经损伤的作用在前期研究中已经得到证实,但详细机制未能明确。同时Notch信号通路和微小RNA系统逐渐成为该领域的潜在热点,但二者的相关作用尚未得到明确报道证实。目的:观察携带骨髓间充质干细胞的小间隙套管对周围神经损伤的治疗作用,探究Notch信号通路和miR-21在治疗过程中的相互作用关系。方法:体内实验中制造SD大鼠喉返神经8 mm缺损模型,分为假手术组、小间隙组和小间隙+骨髓间充质干细胞组,治疗后测定电生理结果,Western blot测定Notch信号通路和微小RNA表达情况。体外实验中在小间隙套管内培养骨髓间充质干细胞并行神经元方向诱导,通过改变Notch/miR 21表达情况,检测神经标记物的表达情况,分析骨髓间充质干细胞活性变化。结果与结论:神经染色和肌电图结果均确认了骨髓间充质干细胞的治疗效果。在骨髓间充质干细胞向神经细胞分化过程中,Notch信号表达明显增强,miR-21表达明显上扬。通过进一步调控两因素表达发现,人为激活Notch信号或使miR-21过表达后骨髓间充质干细胞向神经分化的能力明显增强,抑制Notch信号后,骨髓间充质干细胞神经分化能力减弱,同时miR-21的促进作用失效。发现Notch信号通路和miR-21为骨髓间充质干细胞联合小间隙套接治疗周围神经损伤可能的重要促进机制,其中Notch信号通路具有更高级别的促进或抑制作用等级。BACKGROUND:Nerve injury repair is one of the major surgical hot spots in clinical and basic research.Small-gap sleeve method has been confirmed to have therapeutic effects on peripheral nerve injuries,but the detailed mechanism is unclear.The Notch signaling pathway and the microRNA system have gradually become potential hotspots in this field,but their interactions have not yet been clearly reported.OBJECTIVE:To observe the performance of bone marrow mesenchymal stem cells-packed small-gap sleeve in the treatment of peripheral nerve injuries and to explore the interactions of Notch signaling pathway and miR-21 during the repair process.METHODS:Models of 8 mm-recurrent laryngeal nerve defects were made in Sprague-Dawley rats and randomly divided into three groups:sham operation group,small-gap sleeve group and combined group(small-gap sleeve carrying bone marrow mesenchymal stem cells).After transplantation,neuroelectrophysiological results were measured,and the expression of Notch signal and miR-21 was determined by western blot assay.Bone marrow mesenchymal stem cells cultured in a small-gap sleeve undertook neuronal induction.By changing the expression of Notch/miR21,we detected the expression of neural markers and analyzed the activity of bone marrow mesenchymal stem cells.RESULTS AND CONCLUSION:Transplantation of bone marrow mesenchymal stem cells had therapeutic effects on peripheral nerve injuries as confirmed by neurostaining and electromyogram.During the neural differentiation of bone marrow mesenchymal stem cells,the expressions of Notch signal and miR-21 were significantly increased.Activation of Notch signal or miR-21 overexpression could strengthen the neural differentiation of bone marrow mesenchymal stem cells,while inhibition of Notch signal weakened the neural differentiation of bone marrow mesenchymal stem cells,and miR-21 lost its promotion effect.Overall,Notch signaling pathway and miR-21 are considered to promote the therapeutic effects of bone marrow mesenchymal stem cells combined with smal

关 键 词：喉返神经 周围神经损伤 小间隙套接 MIR-21 NOTCH通路 NICD Hes-1 骨髓基质干细胞 干细胞 受体 Notch 微RNAs 周围神经 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]