中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (19): 3451-3454.doi: 10.3969/j.issn.1673-8225.2011.19.007

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

荧光定量聚合酶链反应检测GATA-1、GATA-2、过氧化物酶体增殖物激活受体γ和干细胞因子基因在再生障碍性贫血患者及正常人骨髓间充质干细胞中的表达

何  丽1，2，肖  扬1，蒋祖军1，李  利1，李勇华1，欧阳玲1，高  飏1，邝丽萍1   

  1. 1解放军广州军区广州总医院血液科，广东省广州市    510010
    2广州中医药大学，广东省广州市  510006
  • 收稿日期:2011-01-04 修回日期:2011-03-02 出版日期:2011-05-07 发布日期:2011-05-07
  • 通讯作者: 肖扬，硕士生导师，主任医师，解放军广州军区广州总医院血液科，广东省广州市 510010 jdxiao111@163.com
  • 作者简介:何丽★，女，1986年生，江西省新余市人，汉族，广州中医药大学在读硕士，主要从事骨髓间充质干细胞对再生障碍性贫血患者造血及免疫调节机制的研究。 heli19860502@qq.com

Real-time PCR detection of GATA-1, GATA-2, peroxisome proliferator-activated receptor gamma, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people

He Li1,2, Xiao Yang1, Jiang Zu-jun1, Li Li1, Li Yong-hua1, Ouyang Ling1, Gao Yang1, Kuang Li-ping1   

  1. 1Hematology Department, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou  510010, Guangdong Province, China
    2Guangzhou University of Traditional Chinese Medicine, Guangzhou  510006, Guangdong Province, China
  • Received:2011-01-04 Revised:2011-03-02 Online:2011-05-07 Published:2011-05-07
  • Contact: Xiao Yang, Master’s supervisor, Chief physician, Hematology Department, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, Guangdong Province, China jdxiao111@163.com
  • About author:He Li★, Studying for master’s degree, Hematology Department, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, Guangdong Province, China; Guangzhou University of Traditional Chinese Medicine, Guangzhou 510006, Guangdong Province, China heli19860502@qq.com

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摘要：

背景：GATA-1、GATA-2、过氧化物酶体增殖物激活受体γ和干细胞因子基因在骨髓造血及成脂调控机制中起重要作用。
目的：观察GATA-1、GATA-2、过氧化物酶体增殖物激活受体γ和干细胞因子基因在再生障碍性贫血患者及正常人间充质干细胞中的表达。
方法：收集6例再生障碍性贫血患者及6位正常人骨髓液各10 mL，Ficoll贴壁法分离培养骨髓单个核细胞，达70%~80%融合后扩增传代。取传至第3代的间充质干细胞，应用流式细胞仪进行鉴定。提取总RNA，比较各组基因与内参基因之间的差别。荧光定量聚合酶链反应检测GATA-1、GATA-2、过氧化物酶体增殖物激活受体γ和干细胞因子基因在再生障碍性贫血患者及正常人间充质干细胞中的表达量。
结果与结论：与正常人间充质干细胞相比，再生障碍性贫血患者间充质干细胞的GATA-2和干细胞因子基因表达量降低(P=0.012，0.039)，过氧化物酶体增殖物激活受体γ基因表达量升高(P=0.035)；两组GATA-1基因表达量差异无显著性意义。提示GATA-2、干细胞因子基因的异常表达可能影响AA患者骨髓微环境的造血调控作用；过氧化物酶体增殖物激活受体γ基因的异常表达可能解释再生障碍性贫血患者骨髓易成脂的原因。

关键词: 再生障碍性贫血, 间充质干细胞, 基因表达, 荧光定量聚合酶链反应, 过氧化物酶增殖物激活爱体&gamma

Abstract:

BACKGROUND: GATA-1, GATA-2, peroxisome proliferator-activated receptor gamma (PPAR-γ), and SCF gene play an important role in bone marrow hematopoietic and concomitant regulation mechanism.
OBJECTIVE: To observe the expressions of GATA-1, GATA-2, PPAR-γ, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people.
METHODS: we collected bone marrow samples from six aplastic anemia patients and six normal persons. Ficoll stick wall method was used to isolate and culture bone marrow mononuclear cells that began to passage at 70%-80% confluence. The 3rd generation of mesenchymal stem cells were detected using flow cytometry. Total RNA was extracted to synthesis and design primer sequence, analysis standard curve, amplification curve, dissolved curve, to be compared with NaCan each gene genetic differences. Fluorescent quantitative PCR detection was employed for GATA-1, GATA-2, PPAR-γ, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people.
RESULTS AND CONCLUSION: Expression of GATA-2 and SCF gene was lower (P=0.012, 0.039) and PPAR-γ expression was higher (P=0.035) in the aplastic anemia patients as compared with normal persons. There was no difference in GATA-1 expression. The findings indicated that abnormal expression of GATA-2 and SCF gene could affect macro-control of bone marrow hematopoietic microenvironment, and abnormal expression of PPAR-γ could explain why bone marrow adipogenesis easily formed in aplastic anemia patients.

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