miR-21对脂肪干细胞移植修复外周神经损伤易发凋亡的干预作用及机制

项目名称： miR-21对脂肪干细胞移植修复外周神经损伤易发凋亡的干预作用及机制

项目编号： No.U1504325

项目类型： 联合基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 张自强

作者单位： 河南科技大学

项目金额： 27万元

中文摘要： 脂肪干细胞是来源于脂肪组织的一种成体干细胞，因其能够分化为雪旺细胞常被用于修复外周神经损伤，然而损伤微环境诱导的细胞凋亡制约了神经再生。微RNA作为重要的调控分子在多种凋亡信号通路中发挥关键作用。申请人前期发现脂肪干细胞凋亡后miR-21显著下调了9.04倍，推测其干预了凋亡过程。由此，本课题拟采用过/抑制miR-21表达的方法，确定miR-21与脂肪干细胞凋亡的关系以及对神经再生的影响。利用生物信息学预测并通过双荧光素酶报告基因系统验证miR-21的靶基因，比较脂肪干细胞凋亡前后该基因的表达，确定miR-21的靶基因。通过生物信息学分析靶基因的信号通路，验证miR-21对凋亡相关信号通路中关键蛋白活化的影响，确定信号通路。单独转染miR-21或共同转染靶基因，检测信号通路中关键蛋白的活化情况，确定miR-21-靶基因-凋亡信号通路轴，揭示miR-21干预脂肪干细胞凋亡发生的分子机制。

中文关键词： 脂肪干细胞；miR-21；神经再生；凋亡

英文摘要： Adipose derive stem cells (ADSCs) isolated from adipose tissue is one kind of adult stem cell, and it is often served as seed cells for treatment of peripheral nerve injury because ADSCs have the ability to differentiated into Schwann cells. However, ADSCs apoptosis induced by microenvironment after injury has been the main obstacle limiting nerve regeneration. MicroRNAs (miRNAs) as important regulatory molecules in cells have been shown to play important roles in a variety apoptotic signal transduction pathway. We found that the expression of miR-21 in apoptotic ADSCs was reduced about 9.04 fold compared to normal ADSCs. Therefore, we think that miR-21 play an important role in the process of ADSCs apoptosis. Thus, the relationship of miR-21 and ADSCs apoptosis, and the effect of miR-21 on nerve regeneration were definited by over expression or suppression expression miR-21. Then we will predict their targets by bioinformatics method, and verify their regulation roles on targets by dual luciferase report gene system. At the same time, the expression of target genes was compared between ADSCs and apoptotic ADSCs. Finally, the targets of miR-21 were definited. Signal transduction pathways of target gene were predicted by bioinformatics method, and effect of miR-21 on activation of main proteins in apoptotic signal transduction pathways were evaluated. Then we could definite signal transduction pathways of miR-21 in the process of ADSCs apoptosis. ADSCs were transfected with miR-21 alone or together with target gene, and activation of main proteins in apoptotic signal transduction pathways were evaluated. Finally, we could definite miR-21-target gene-apoptotic signal transduction pathway according to above results, and finally reveal molecule mechanism of miR-21 to interfere in the process of ADSCs apoptosis.

英文关键词： Adipose derive stem cell;miR-21;Nerve regeneration;Apoptosis