miR-17-5p在不同代骨髓间充质干细胞中的表达

Expression of miR-17-5p in bone marrow mesenchymal stem cells in different generations

目的:观测miR-17-5p在年轻的和衰老骨髓间充质干细胞(BMMSCs)中的表达,并分析其与细胞衰老的相关性。方法:采用密度梯度离心法分离大鼠BMMSCs并培养至第10代。取第4代(P4)BMMSCs和第10代(P10)BMMSCs的用细胞衰老β-半乳糖苷酶(β-gal)染色法进行染色。采用实时定量PCR检测P4及P10 BMMSCs中miR-17-5p基因的表达,并用2(-ΔΔct)法定量分析。结果:①形态学改变:P4～P10的细胞形态发生改变,P4的细胞形态均一,呈典型的漩涡状生长,P10的细胞失去典型的梭形外观,细胞体积增大而不规则,形状扁平破碎,细胞间隙增大,出现凋亡。②β-gal染色结果:P10的细胞肥大、染呈蓝色(呈阳性),P4的细胞染色后未着色(呈阴性)。③实时定量PCR检测结果:P4细胞miR-17-5p基因的表达量是P10细胞的6.5倍(P<0.01)。结论:衰老BMMSCs中miR-17-5p的表达量显著降低,提示miR-17-5p可能参与了细胞的衰老过程。

AIM： To detect the expression of miR-17-5p between young and aging bone marrow mesenchymal stem cells（BM-MSCs） and to analyze its correlation with cellular senescence.METHODS： BM-MSCs were isolated from bone marrow of Sprague Dawley rats by density gradient centrifugation and cultivated in vitro to the 10th generation.The fourth generation BM-MSCs（P4） and 10th generation BM-MSCs（P10） were stained with β-gal staining.2（-ΔΔct） method was used for quantitative analysis of expression of miR-17-5p between young and aging BM-MSCs by real-time RT-PCR.RESULTS： Morphological changes： P4 to P10 showed cell morphology changes.The shape of P4 cells was highly homogeneous and showed typical swirling growth.P10 cells lost the typical fusiform,increased irregularly in cell volume and were flat and broken in shape.The cell gap increased and showed apoptosis.β-Gal staining results： P10 cells were hypertrophic,blue-stained and positive.P4 cells were not colored after being stained and exhibited negative results.Real-time PCR results： the expression of miR-17-5p in P4 was 6.5 times that of P10（P〈0.01）.CONCLUSION： Aging BM-MSCs miR-17-5p expression levels are significantly lower,suggesting that miR-17-5p may be involved in the process of cell aging.