miR-186-5p修饰骨髓间充质干细胞通过下调FZD3对脊髓损伤大鼠具有神经保护作用  

Bone marrow mesenchymal stem cells overexpressed with miR-186-5p have a neuroprotective effect on rats with spinal cord injury by down-regulating FZD3

作　　者：季英楠[1] 东黎阳[1] 冯时 JI Ying-nan;DONG Li-yang;FENG Shi(Department of Orthopedics,Affiliated Central Hospital of Shenyang Medical College,Shenyang 110024,China)

机构地区：[1]沈阳医学院附属中心医院骨科,辽宁沈阳110024

出　　处：《解剖科学进展》2021年第5期549-553,共5页Progress of Anatomical Sciences

基　　金：沈阳医学院科技基金项目(20142035)。

摘　　要：目的探讨miR-186-5p修饰的骨髓间充质干细胞对脊髓损伤大鼠的神经保护作用及可能机制。方法 40只SD大鼠随机分为假手术组(Sham组)、脊髓损伤模型组(SCI组)、骨髓间充质干细胞移植组(BMSCs组)和miR-186-5p修饰的骨髓间充质干细胞移植组(miR+BMSCs组),每组10只;除了Sham组,其余三组通过Allen’s法建立SCI大鼠模型,移植后第1、7、14、21天对各组大鼠进行运动功能(BBB)评分;HE染色观察各组大鼠脊髓损伤病理学变化;IF检测NeuN表达情况;ELISA检测NT-3含量变化;利用qRT-PCR检测各组大鼠miR-186-5p的表达;荧光素酶实验验证miR-186-5p与FZD3的关系;Western blot方法检测损伤脊髓组织中的FZD3、β-catenin、c-Myc、cyclinD1的表达水平变化。结果与SCI组相比,BMSCs组和miR+BMSCs组的BBB评分、损伤脊髓组织中NeuN的表达、NT-3的表达以及β-catenin、c-Myc、cyclinD1的蛋白表达均明显升高(P<0.05),FZD3的蛋白表达明显下降(P<0.05)。FZD3被预测并确认为miR-186-5p的靶基因。与BMSCs组相比,miR+BMSCs组的BBB评分、损伤脊髓组织中NeuN的表达、NT-3的表达以及β-catenin、c-Myc、cyclinD1的蛋白表达显著升高(P<0.05),FZD3的蛋白表达呈下降趋势(P<0.05)。结论 miR-186-5p修饰的骨髓间充质干细胞移植可通过下调FZD3对SCI大鼠的神经提供保护作用。Objective To explore the neuroprotective effect of miR-186-5 p modified bone marrow mesenchymal stem cells(BMSCs) on spinal cord injury rats. Methods 40 SD rats were randomly divided into sham operation group(Sham group), spinal cord injury model group(SCI group), bone marrow mesenchymal stem cells(BMSCs) treatment group and miR-186-5 p modified BMSCs treatment group(miR+BMSCs group), 10 rats in each group. SCI rat model was established by Allen’s method. Rats in each group were scored for motor function(BBB). HE staining were used to observe the pathological changes of the spinal cord. ELISA was used to detect the content of NT-3. The expression of NeuN was detected by IF. The level of miR-186-5 p was analyzed by RT-qPCR. The relationship between miR-186-5 p and FZD3 was confirmed by luciferase assay. The expression of FZD3, β-catenin, c-myc and cyclinD1 in injured spinal cord were detected by Western blot. Results Compared with SCI group, BBB score, expression of NeuN and content of NT-3 of injured spinal cord tissue were increased in BMSCs group and miR+BMSCs group. FZD3 was predicted and confirmed as a target gene of miR-186-5 p. Compared with BMSCs group, BBB score, the expression of NeuN, the expression of NT-3, β-catenin, c-myc and CyclinD1 of injured spinal cord tissue was increased significantly(P<0.05), and the expression of FZD3 was decreased(P<0.05). Conclusion miR-186-5 p modified bone marrow mesenchymal stem cell transplantation can protect the nerves of SCI rats by down regulating FZD3.

关 键 词：miR-186-5p 骨髓间充质干细胞 脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]