中国全科医学2017，Vol.20Issue(33)：4156-4161,6.DOI:10.3969/j.issn.1007-9572.2017.00.122
Let-7b对骨髓基质干细胞诱导分化为神经元的作用研究
Effect of Let-7b on the Differentiation of Bone Marrow Mesenchymal Stem Cells into Nerve Cells
摘要
Abstract
Objective To investigate the role of Let-7b in the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into neurons.Methods This study was conducted in 2013.Eighty SPF grade SD rats were randomly divided into blank control group,negative control group,Let-7b-up lentiviral vector (LV-mo-Let-7b-up) group,Let-7b-inhibition lentiviral vector (LV-rno-Let-7b-inhibition) group,20 rats in each group.The negative control group,LV-mo -Let-7b-up group,LV-rno-Let-7b-inhibition group were injected EGFP blank virus respectively.Four groups of rats were sacrificed by cervical dislocation,and MSCs were obtained and cultured for 4 generations.Subculture of MSCs were infected with lentivirus,that is,the blank control group and the negative control group were not infected,LV-rno-Let-7b-up group,LV-rno-Let-7b-inhibition group were infected with LV-rno-Let-7b-up,Let-7b-inhibition,respectively.The survival rate of MSCs was detected by MTT before infection,at 24 h after infection,on the fifth day after infection.Four groups were induced by fasudil,and the differentiation efficiency of MSCs into neurons was observed.The levels of NSE,MAP-2 and Tau were detected by immunohistochemistry.The levels of NSE mRNA,MAP-2 mRNA and Tau mRNA were detected by RT-PCR.Results The MSCs of LV-rno-Let-7b-up group had the strongest fluorescence expression on the third day after LV-rno-Let-7b-up infection.The MSCs of LV-rno-Let-7b-inhibition group had the strongest fluorescent expression on the fifth day after LV-rno-7b-inhibition infection.On the fifth day after infection,the MSCs infection rates of the negative control group,LV-rno-Let-7b-up group and LV-rno-Let-7b-inhibition group were 87.46％,86.81％ and 87.12％,respectively.At 24 h after infection,and on the fifth day after infection the blank control group presented higher survival rate of MSCs than the other three groups (P ＜ 0.05).The survival rate of MSCs on the fifth day after infection was found to be higher than that at 24 h after infection in the negative control group,LV-rno-Let-7b-up group and LV-mo-Let-7b-inhibition group (P ＜ 0.05).Some MSCs showed neuronal changes in the negative control group and the blank control group,almost all the MSCs showed typical neuron-like structures in LV-rno-Let-7b-up group,but only a small number of MSCs showed a typical neuron-like structure in the LV-rno-Let-7b-inhibition group.The protein and mRNA levels of NSE,MAP-2,Tau in LV-rno-Let-7b-up group were higher than those in blank control group and negative control group (P ＜ 0.05);the protein and mRNA levels of NSE,MAP-2,Tau in LV-rno-Let-7b-inhibition group were lower than those in blank control group and negative control group (P ＜ 0.05).Conclusion We found that the differentiation efficiency of MSCs into neurons was increased after infection with LV-rno-Let-7b-up,which suggests that Let -7b promotes the differentiation of MSCs into neurons.关键词
神经元/干细胞,骨髓/慢病毒属/Let-7b/细胞分化Key words
Neurons/Stem cells, myeloid/Lentivirus/Let-7b/Cell differentiation分类
医药卫生引用本文复制引用
李进领,李智,李琳,张静,贾延劼,安颖..Let-7b对骨髓基质干细胞诱导分化为神经元的作用研究[J].中国全科医学,2017,20(33):4156-4161,6.基金项目
国家自然科学基金资助项目(81071114,81371385) （81071114,81371385）
