HIF-1α induced-VEGF overexpression in bone marrow stem cells protects cardiomyocytes against ischemia

Ying Dai, Meifeng Xu, Yigang Wang, Zeeshan Pasha, Tingyu Li, Muhammad Ashraf

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

Hypoxia inducible factor-1α (HIF-1α) is a proangiogenic transcription factor stabilized and activated under hypoxia. It regulates the expression of numerous target genes, including vascular endothelial growth factor (VEGF) and other cytoprotective proteins. In this study, we hypothesized that bone marrow stem cells (BMSCs) secrete growth factors which protect cardiomyocytes via HIF-1α pathway. BMSCs were obtained from transgenic mice overexpressing green fluorescent protein (GFP). The study was carried out in vitro using co-culture of BMSCs with cardiomyocytes. LDH release, MTT uptake, DNA fragmentation and annexin-V positive cells were used as cell injury markers. The level of HIF-1α protein as well as its activated form and VEGF were measured by ELISA. The expression of HIF-1α and VEGF in BMSCs was analyzed by quantitative PCR and cellular localization was determined by immunohistochemistry. LDH release was increased and MTT uptake was decreased after exposure of cardiomyocytes to hypoxia for 30 h, which were prevented by co-culturing cardiomyocytes with BMSCs. Cardiomyocyte apoptosis induced by hypoxia and H2O2 was also reduced by co-culture with BMSCs. VEGF release from BMSCs was significantly increased in parallel with high level of HIF-1α in BMSCs following anoxia or hypoxia in a time-dependent manner. Although no significant up-regulation could be seen in HIF-1α mRNA, HIF-1α protein and its activated form were markedly increased and translocated to the nucleus or peri-nuclear area. The increase and translocation of HIF-1α in BMSCs were completely blocked by 2-methoxyestradiol (2-ME2; 5 μmol), a HIF-1α inhibitor. Moreover, the protection of cardiomyocytes by BMSC and VEGF secretion was abolished by neutralizing HIF-1α antibody in a concentration dependent manner (200-3200 ng/ml). Bone marrow stem cells protect cardiomyocytes by up-regulation of VEGF via activating HIF-1α.

Original languageEnglish (US)
Pages (from-to)1036-1044
Number of pages9
JournalJournal of molecular and cellular cardiology
Volume42
Issue number6
DOIs
StatePublished - Jun 1 2007
Externally publishedYes

Fingerprint

Hypoxia-Inducible Factor 1
Cardiac Myocytes
Bone Marrow Cells
Vascular Endothelial Growth Factor A
Stem Cells
Ischemia
Cell Hypoxia
Coculture Techniques
Up-Regulation
Proteins
Annexin A5
DNA Fragmentation
Green Fluorescent Proteins
Transgenic Mice
Intercellular Signaling Peptides and Proteins
Transcription Factors
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry

Keywords

  • Bone marrow stem cells
  • Cardiomyocytes
  • HIF-1α
  • HIF-1α neutralizing antibody
  • Hypoxia
  • VEGF

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

HIF-1α induced-VEGF overexpression in bone marrow stem cells protects cardiomyocytes against ischemia. / Dai, Ying; Xu, Meifeng; Wang, Yigang; Pasha, Zeeshan; Li, Tingyu; Ashraf, Muhammad.

In: Journal of molecular and cellular cardiology, Vol. 42, No. 6, 01.06.2007, p. 1036-1044.

Research output: Contribution to journalArticle

Dai, Ying ; Xu, Meifeng ; Wang, Yigang ; Pasha, Zeeshan ; Li, Tingyu ; Ashraf, Muhammad. / HIF-1α induced-VEGF overexpression in bone marrow stem cells protects cardiomyocytes against ischemia. In: Journal of molecular and cellular cardiology. 2007 ; Vol. 42, No. 6. pp. 1036-1044.
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AU - Ashraf, Muhammad

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