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|本期目录/Table of Contents|

miR-17-5p在不同代骨髓间充质干细胞中的表达

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2013年第3期
页码:
333-336
栏目:
基础研究
出版日期:
2013-06-25

文章信息/Info

Title:
Expression of miR-17-5p in bone marrow messenchymal stem cells in different generations
作者:
达 晶; 吕安林; 侯兆蕾; 侯 红; 艾世宜; 侯 婧; 刘博武
(第四军医大学西京医院心血管内科，陕西 西安 710032)
Author(s):
DA Jing;  LV An lin;  HOU Zhao lei;  HOU Hong;  AI Shi yi;  HOU Jing;  LIU Bo wu
(Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China)
关键词:
骨髓间充质干细胞; miR-17-5p; 细胞衰老; 实时定量PCR
Keywords:
bone marrow mesenchymal stem cells;  miR-17-5p;  cellular senescence;  realtime PCR
分类号:
R339.38
DOI:
-
文献标识码:
A
摘要:
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。
Abstract:
AIM:To detect the expression of miR-17-5p between young and aging bone marrow mesenchymal stem cells (BMMSCs) and to analyze its correlation with cellular senescence. METHODS: BMMSCs were isolated from bone marrow of SD rats by density gradient centrifugation and cultivated in vitro to the 10th generation. The fourth generation BMMSCs (P4) and 10th generation BMMSCs (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 bone marrow mesenchymal stem cells (BMMSCs) 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. The 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 BMMSCs miR-17-5p expression levels are significantly lower, suggesting that the miR-17-5p may be involved in the process of cell aging.

参考文献/References

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备注/Memo

备注/Memo:
收稿日期：2013-01-22.
通讯作者：吕安林，主任医师，主要从事冠心病和干细胞移植研究 Email：lvanlin@fmmu.edu.cn
作者简介：达晶，硕士生Email：crystalda118@yahoo.cn
更新日期/Last Update: 2013-07-16