微小RNA-124在骨髓间充质干细胞分化为神经源性细胞的作用及其对脊髓损伤修复的影响  

Effect of microRNA - 124 on the differentiation of bone marrow mesenchymal stem cells into neurogenic cells and its effect on spinal cord injury repair

作　　者：路坦[1] 尉娜[2] 张超[1] 董玉珍[1] 赵斌[1] 

机构地区：[1]新乡医学院第一附属医院骨外二科,河南卫辉453100 [2]新乡医学院第三附属医院神经内科,河南卫辉453003

出　　处：《中华实验外科杂志》2016年第1期179-181,共3页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划（豫卫科201304029）

摘　　要：目的探讨微小RNA-124（miR-124）过表达对骨髓间充质干细胞（BMSCs）分化为神经元样细胞的作用、分子机制及神经元样细胞对脊髓损伤修复的影响。方法通过从小鼠骨髓中分离得到BMSCs，pre—miR-124转染BMSCs，利用实时定量反转录聚合酶链反应（RT—qPCR）检测miR-124的表达量，Western blot检测多聚嘧啶序列结合蛋白1（PTBP1）、多聚嘧啶序列结合蛋白2（PTBP2）的表达。BMSCs体外诱导分化为神经元样细胞，移植入损伤脊髓中。结果BMSCs在特定条件的培养基下能分化为神经元样细胞，当miR-124过表达时，能提高其分化效率。PTBP1对PTBP2的表达有阻遏作用，神经源性分化的过程中，miR-124的过表达抑制PTBP1的表达，从而增加PTBP2的表达量，提高神经元样分化效率。miR-124过表达的BMSCs分化的神经元样细胞移植入损伤脊髓，能加速损伤脊髓修复。pre-miR-124转染的BMMSCs分化为神经源性细胞的分化率显著高于未转染的骨髓间充质干细胞，对应的标记基因神经元特异性烯醇化酶（NSE）、β-微管蛋白Ⅲ及胶质纤维酸性蛋白（GFAP）的表达呈阳性的细胞比例分别为50％、58％和30％，与对照组（17％、24％和20％）比较差异有统计学意义（P〈0．05）。pre-miR-124转染的BMMSCs分化为神经源性细胞的分化率显著高于未转染的BMMSCs。结论miR-124能促进BMSCs分化为神经元样细胞，神经元样细胞移植入受损脊髓中能加速其修复。Objective To make clear the effect and molecular mechanism of microRNA - 124 （ miR - 124） over - expression on the differentiation of bone marrow mesenchymal stem cells （BMSCs） into neurogenic cells and the role of neurogenic cells on the repair of spinal cord injury. Methods BMSCs were isolated from bone marrow and transfected with adenoviruses for pre - miR - 124, real- time reverse transeriptase- polymerase chain reaction（ RT- qPCR） was used to detect the expression of miR- 124. Western blotting was used to detect the expressions of polypyrimidine tract - binding protein 1 （ PTBP1 ） and polypyrimidine tract- binding protein 1 （PTBP2）. The neurogenic cells from BMSCs were transplanted into model of spinal cord injury. Results MiR - 124 over - expression promoted BMSCs differentiation into neurogenic cells in vitro. MiR - 124 reduced the expression of PTBP1 by targeting PTBP1 at the 3＇ untranslated region （3＇ -UTR）. The protein level of PTBP2 was up -regulated when down- regulation of PTBP1. PTBP2 increased the productivity of BMSCs differentiation into neurogenic cells. Neurogenic cells from BMSCs of miR - 124 over - expression were used to transplant and take part in tissue regeneration in spinal cord injury. Between the pre - miR - 124 transfeetion of BMSCs differentiation for neurogenic differentiation rate was significantly higher than without transfection between BMSCs. The corresponding marker genes neuron specific enolase （NSE）, beta tubulin 111 and glial fibrillary acidic protein （GFAP） expression levels with transfeetion by pre - miR - 124 of BMSCs were 50% , 58% and 30% respectively, that significantly higher than that of without transfection of BMSCs were 17% ,24% and 20% respectively （P 〈0. 05 ）. Conclusion This study show that miR - 124 promotes BMSCs differentiation into neurogenic cells in vitro. BMSCs - derived neurogenic cells were transplanted into injured spinal for transplantation, and successfully participated in neural restoration. It can be a new

关 键 词：微小RNA 骨髓 间充质干细胞 神经源性细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]