中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6759-6763.doi: 10.3969/j.issn.1673-8225.2010.36.024

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

缺氧诱导因子1α小干扰RNA对骨髓间充质干细胞缺氧诱导因子1α、基质细胞衍生因子1α和血管内皮生长因子基因表达的影响

张文文1，林  明1，许昌声2   

  1. 1福建医科大学附属第一临床学院心内科，福建省福州市  350005；  2福建省高血压研究所，福建省福州市  350005
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 通讯作者: 林明，主任医师，教授，福建医科大学附属第一医院心内科，福建省福州市 350005 linming0039@126.com
  • 作者简介:张文文★，女， 1983年生，陕西省宝鸡市人，汉族，福建医科大学在读硕士，医师，主要从事干细胞方面的研究。 xw132202@163.com
  • 基金资助:

    福建医科大学教授学术发展基金项目（JS06040）“肝细胞生长因子转染骨髓间充质干细胞移植治疗心肌病心力衰竭”。

Effects of hypoxia inducible factor-1 alpha small interfering RNA on hypoxia inducible factor-1 alpha, stromal derived factor 1 alpha and vascular endothelial growth factor gene expression in bone marrow mesenchymal stem cells 

Zhang Wen-wen1, Lin Ming1, Xu Chang-sheng2   

  1. 1 Department of Cardiology, First Clinical College Affiliated to Fujian Medical University, Fuzhou  350005, Fujian Province, China; 2 Fujian Hypertension In stitute, Fuzhou  350005, Fujian Province, China
  • Online:2010-09-03 Published:2010-09-03
  • Contact: Lin Ming, Chief physician, Professor, Department of Cardiology, First Clinical College Affiliated to Fujian Medical University, Fuzhou 350005, Fujian Province, China linming0039@126. com
  • About author:Zhang Wen-wen, Studying for master’s degree, Physician, Department of Cardiology, First Clinical College Affiliated to Fujian Medical University, Fuzhou 350005, Fujian Province, China xw132202@163.com
  • Supported by:

    the Academic Development Foundation for Professor in the Fujian Medical University, No. JS06040

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摘要：

背景：缺氧可以影响骨髓间充质干细胞分泌细胞因子，其机制可能是通过缺氧诱导因子1α起作用的。
目的：观察缺氧诱导因子1α RNA干扰对骨髓间充质干细胞缺氧诱导因子1α、基质细胞衍生因子1α和血管内皮生长因子基因表达的影响。
方法：贴壁法培养骨髓间充质干细胞，取第3~5代细胞用于实验，倒置显微镜下观察细胞形态，并用流式细胞仪检测其表面标志物CD34、CD44、CD90表达。将骨髓间充质干细胞分四组：常氧对照组(无干预因素)、缺氧组(缺氧24 h)、脂质体对照组(转染空载脂质体后缺氧24 h)、RNA干扰组(转染脂质体介导的RNA干扰序列后缺氧24 h)。RT-PCR法检测骨髓间充质干细胞的缺氧诱导因子1α、基质细胞衍生因子1α和血管内皮生长因子 mRNA表达水平， ELISA检测骨髓间充质干细胞培养上清缺氧诱导因子1α、基质细胞衍生因子1α和血管内皮生长因子蛋白表达水平。
结果与结论：同常氧对照组比较，缺氧组缺氧诱导因子1α、基质细胞衍生因子1 α和血管内皮生长因子基因及蛋白表达增高(P < 0.05)；同脂质体对照组比较，RNA干扰组缺氧诱导因子1α、基质细胞衍生因子1α和血管内皮生长因子基因及蛋白表达降低(P < 0.05)。证实了缺氧条件可以使骨髓间充质干细胞血管内皮生长因子和基质细胞衍生因子1α的表达增加，抑制缺氧诱导因子1α的表达可以使血管内皮生长因子和基质细胞衍生因子1α的表达减少，缺氧诱导因子1α很可能是干细胞移植细胞因子分泌的调控因素。

关键词: 基质细胞衍生因子, 骨髓间充质干细胞, RNA干扰, 缺氧诱导因子, 血管内皮生长因子

Abstract:

BACKGROUND: The cytokines secreted by bone marrow mesenchymal stem cells (BMSCs) can be affected by hypoxia. Its mechanism may exert effects via hypoxia inducible factor-1 α (HIF-1 α).
OBJECTIVE: To observe HIF-1 α, stromal derived factor 1 α (SDF-1 α) and vascular endothelial growth factor (VEGF) gene expression affected by HIF-1 α small interfering RNA (siRNA) in BMSCs.
METHODS: BMSCs were cultured by adherence method. The third to fifth passages of BMSCs were selected for experiments. Inverted microscope was used to observed morphology. Flow cytometry was used to detect the expression of surface markers CD34, CD44 and CD90. BMSCs were divided into four groups, normal control group (without any treatment), hypoxia group (hypoxia for 24 hours), liposome control group (BMSCs transfected with liposome then hypoxia for 24 hours), RNA interference (BMSCs transfected with RNA interference sequences then hypoxia for 24 hours). Reverse transcription-polymerase chain reaction was utilized to determine HIF-1 α, SDF-1 α and VEGF mRNA expression in BMSCs. Enzyme linked immunosorbent assay was employed to detect HIF-1 α, SDF-1 α and VEGF protein expression in BMSCs.
RESULTS AND CONCLUSION: Compared with the normal control group, HIF-1 α, SDF-1 α and VEGF mRNA and protein expression increased in the hypoxia group (P < 0.05). Compared with the lipsome control group, HIF-1 α, SDF-1 α and VEGF mRNA and protein expression decreased in the RNA interference group (P < 0.05). These verified that hypoxia increased SDF-1 and VEGF gene expression. Inhibition of HIF-1 α expression decreased SDF-1 α and VEGF expression. HIF-1 α may be a regulatory factor for cytokine secretion following stem cell transplantation.

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