科研之友
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摘要
Hypoxia inducible factor-1 alpha (HIF-1 alpha) 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-1 alpha 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-1 alpha protein as well as its activated form and VEGF were measured by ELISA. The expression of HIF-1 alpha 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-1 alpha in BMSCs following anoxia or hypoxia in a time-dependent manner. Although no significant up-regulation could be seen in HIF-1 alpha mRNA, HIF-1 alpha protein and its activated form were markedly increased and translocated to the nucleus or peri-nuclear area. The increase and translocation of HIF-1 alpha in BMSCs were completely blocked by 2-methoxyestradiol (2-ME2; 5 mu mol), a HIF-1 alpha inhibitor. Moreover, the protection of cardiomyocytes by BMSC and VEGF secretion was abolished by neutralizing HIF-1 alpha antibody in a concentration dependent manner (200-3200 ng/ml). Bone marrow stem cells protect cardiomyocytes by up-regulation of VEGF via activating HIF-1 alpha.
- 出版日期2007-6
- 单位重庆医科大学
