中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (27): 4991-4995.doi: 10.3969/j.issn.2095-4344.2012.27.010

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

5-氮胞苷诱导兔脂肪干细胞分化为心肌样细胞的特定基因表达

宋艳琴，张艳梅，王妮妮，修 金，张瑞萍   

  1. 辽河油田总医院循环内科，辽宁省盘锦市 124000
  • 收稿日期:2012-02-23 修回日期:2012-06-03 出版日期:2012-07-01 发布日期:2013-11-01
  • 作者简介:宋艳琴★，女，1974年生，辽宁省辽中县人，汉族，2008年中国医科大学毕业，硕士，主治医师，主要从事心血管疾病诊断与治疗方面的研究。 2545150599@qq.com

Cardiac-specific gene expression in cardiomyocytes differentiated by rabbit adipose-derived stem cells under 5-azacytidine induction

Song Yan-qin, Zhang Yan-mei, Wang Ni-ni, Xiu Jin, Zhang Rui-ping   

  1. Department of Cardiology, General Hospital of Liaohe Oil Field, Panjin 124000, Liaoning Province, China
  • Received:2012-02-23 Revised:2012-06-03 Online:2012-07-01 Published:2013-11-01

PDF

309

被引次数

11

摘要：

背景：采用生物学技术将脂肪干细胞进行心肌内移植，修复损伤心肌组织是近年来心血管病研究领域的一大热点。脂肪干细胞经诱导分化后的特定心肌基因表达尚无全面的研究报道。
目的：观察兔脂肪干细胞经5-氮胞苷诱导分化为心肌样细胞后心肌特异性基因的表达。
方法：取第2代脂肪干细胞，加入2.5，5，10，20，40，80 μmol/L 5-氮胞苷经24 h作用后，用完全培养液对其继续进行培养，4周后再观察细胞生长和形态学的改变。待诱导1，2，3，4周，利用RT-PCR技术检测心钠素，脑钠素，α-骨骼肌动蛋白的基因表达。
结果与结论：5-氮胞苷诱导1周，其细胞大多呈紧密平行排列，其体积有所增大，梭形细胞比例相对有所下降，40，80 μmol/L组细胞形态变化明显。诱导第4周时，40，80 μmol/L组细胞折光性减弱，细胞活性减弱，细胞死亡数继续增加。RT-PCR检测心钠素，脑钠素，α-骨骼肌动蛋白的基因表达呈逐渐增加趋势；10 μmol/L组各基因的表达明显高于其他各组(P < 0.05)。结果提示兔脂肪干细胞经5-氮胞苷诱导分化的细胞具有心肌细胞的某些基因的特异性表达。

关键词: 心肌样细胞, 心钠素, 脂肪干细胞, 脑钠素, α-骨骼肌动蛋白, 基因表达, 5-氮胞苷, 干细胞

Abstract:

BACKGROUND: Intramyocardial transplantation of adipose-derived stem cells using biological technique for repair of injured myocardial tissue is a gradually increasing research field of cardiovascular disease. There has been no report describing cardiac-specific gene expression in cardiomyocytes induce-differentiated by rabbit adipose-derived stem cells.
OBJECTIVE: To investigate cardiac-specific gene expression in cardiomyocytes differentiated by rabbit adipose-derived stem cells under 5-azacytidine induction.
METHODS: Passage 2 adipose-derived stem cells were treated with 2.5, 5, 10, 20, 40, 80 μmo1/L 5- azacytidine for 24 hours and cultured with complete medium without 5-azacytidine. Four weeks later, cellular growth and morphological changes were observed. At the first, second, third and fourth weeks, the mRNA expression of atrial natriuretic peptide, brain natriuretic peptide and α-skeletal actin in adipose-derived stem cells was detected by RT-PCT.
RESULTS AND CONCLUSION: After 5-azacytidine induction for 1 week, the majority of cells were arranged in parallel, with increased soma, and the proportion of shuttle-shaped cells was decreased, in particular in the 40, 80 μmol/L 5- azacytidine groups. At the fourth week, cell refraction was attenuated, cell activity was weakened, and more cells died in the 40, 80 μmol/L 5-azacytidine groups. The mRNA expression of atrial natriuretic peptide, brain natriuretic peptide and α-skeletal actin in adipose-derived stem cells increased gradually. The mRNA expression of atrial natriuretic peptide, brain natriuretic peptide and α-skeletal actin in the 10 μmol/L 5-azacytidine group was significantly higher than that in the other groups (P < 0.05). 5-azacytidine induced rabbit adipose-derived stem cells show cardiac-specific gene expression.

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