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5-氮杂胞苷诱导大鼠骨髓间充质干细胞分化为心肌细胞的实验研究(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2004年第6期
页码:
505-508,512
栏目:
基础研究
出版日期:
2004-11-01

文章信息/Info

Title:
Experimental study on cardiomyocytes induced by 5-Azacytidine from mouse marrow stromal cells
作者:
李　雪;  贾国良;  刘　兵; 秦　涛
第四军医大学西京医院心内科,陕西西安710032
Author(s):
LI Xue ;  JIA Guo-liang ;  LIU Bing ;  QIN Tao
Department of Cardiology , Xijing Hospital , Fourth Military MEdical Vniversity , Xi’an , Shaanxi 710033 ,China
关键词:
骨髓间充质干细胞; 分化; 心肌样细胞
Keywords:
marrow st romal cell ;  differentiation ;  cardiomyocyte
分类号:
R331. 31
DOI:
-
文献标识码:
A
摘要:
目的:观察5-氮杂胞苷(5-Azacytidine) 能否诱导骨髓间充质干细胞分化为心肌样细胞及其分化率,并探明骨 髓间充质干细胞分化为心肌样细胞外,尚有哪些细胞类型存在? 方法:以成年SD 大鼠为研究对象,分离单个核细 胞,用5-氮杂胞苷诱导骨髓间充质干细胞的分化。通过倒置显微镜观察受到诱导的骨髓间充质干细胞形态的变 化,用免疫组化的方法标测分化的心肌样细胞的肌钙蛋白I (troponin I) 和结蛋白(desmin) 是否存在? 并用放免试剂 盒测定分化的心肌样细胞能否分泌心房钠尿肽,透射电镜可以观察到分化的心肌样细胞内是否有肌丝和肌丝团的 存在,RT-PCR 方法可以测定分化的心肌样细胞是否有Nkx2. 5 和troponin I 的cDNA 的表达? 从不同的层面鉴定 诱导分化的心肌样细胞是否具有心肌细胞的特性和功能? 并用流式细胞仪测定分化的心肌样细胞占细胞总数的 百分比。结果: ①5-氮杂胞苷可以诱导骨髓间充质干细胞分化为心肌样细胞。②在骨髓间充质干细胞分化为心肌 样细胞的过程中,有部分成纤维细胞生长和未分化的单个核细胞存在。结论:5-氮杂胞苷可以诱导骨髓间充质干细 胞分化为心肌样细胞,5-氮杂胞苷诱导骨髓间充质干细胞分化为心肌样细胞后再做细胞移植,可能有改善受损心 脏的功能。
Abstract:
AIM:Marrow st romal cells were isolated f row SD mice and were then induced by 5-azacytidine to confirm whether 5-azacytidine can induced marrow st romal cells into cardiomyocytes and the differentiation rate and to make sure of the cells involved in the course besides cardiomyocytes. METHODS : We use 2 month male SD mice were used as our subject s. Marrow st romal cells were obtoined by proportion 1.073 percoll. 24 hours later marrow st romal cells were t reated with 5-azacytidine . Marrow st romal cells show round morphology and there is a single nucleus before t reated. The morphology changed after the cells were t reated with 5-azacytidine. They prot ruded pseudopods and connected with adjoining cells in one week and they connected together in 2～3 weeks. They grow clusterece forming muscle islands in four weeks. They secrectea at rial riat riuretic peptide. They were staina with anti-troponin-I and anti-desnim antinodies. Elect ron microscopy reveuled myotubes st ructure ,and there were bridge-granules st ructures bet reen adjoin cells. Tthese cells expressed Nkx2. 5 and t roponin I cDNA cardiomyocytes defferentiation was examined by flow cytometer . This cell line provided a powerful model for the study of cardiomyocytes differentiation. RESULTS : ① 5-acytidine could induce bone marrow st romal cell into cardiomyocytes and the differentiation rate was about 25 % . ②During marrow st romal cell differentiation , there was fibroblast cells and some round single nueclea cells.CONCLUSION : Our study shows that 52azacytidine can induce bone marrow st romal cells into cardiomyocytes. After marrow st romal cells are t reated with 52azacytidine and grafted , they can improve damaged heart function.

参考文献/References

[1] 张航向,李　源,臧益民. 干细胞移植与心血管疾病[J ] . 心脏杂志,2002 ,14 (6) :524 - 529.

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[ 5 ] Makino S , Fukuda K,Miyoshi S , et al . Cardiomyocytes can be generated from marrow stromal cells i n vit ro[ J ]. J Cli n Invest ,1999 , 103 (5) :697 - 705.

[6 ] 李　雪,贾国良,王海昌,等. 成体干细胞在心血管系统疾病中的应用[A] . 朱妙章,袁文俊,吴博威,等. 心血管生理学与临床[M] . 北京:高等教育出版社,2004. 439 - 444.

备注/Memo

备注/Memo:
收稿日期：2004-5-3
更新日期/Last Update: 2004-11-01