Hypoxia inducible factor-1α mediates protective effects of ischemic preconditioning on ECV-304 endothelial cells

• Published in: World journal of gastroenterology : WJG Vol. 13; no. 16; pp. 2369 - 2373
• Main Author: Shi, Liu-Bin
• Format: Journal Article
• Language: English
• Published: Department of Organ Transplantation, Huashan Hospital, Fudan University, Shanghai 200040, China%Institute of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China%Department of General Surgery, Ruijin Hospital,School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China 28.04.2007; Baishideng Publishing Group Co., Limited
• Subjects: Rapid Communication; Anoxia/reoxygenation injury; Hypoxia inducible factor-1α; Preconditioning; Reperfusion injury; Endothelial cells
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v13.i16.2369

R3; AIM: To investigate whether hypoxia inducible factor-1α (HIF-1α) is linked to the protective effects of ischemic preconditioning (IP) on sinusoidal endothelial cells against ischemia/reperfusion injury.METHODS: Sinusoidal endothelial cell lines ECV-304 were cultured and divided into four groups: control group, cells were cultured in complete DMEM medium; cold anoxia/warm reoxygenation (A/R) group, cells were preserved in a 4℃ UW solution in a mixture of 95% N2 and 5% CO2 for 24 h; anoxia-preconditioning (ARC) group, cells were treated with 4 cycles of short anoxia and reoxygenation before prolonged anoxia-preconditioning treatment; and anoxia-preconditioning and hypoxia inducible factor-1α (HIF-1α) inhibitor (I-HIF-1) group, cells were pretreated with 5 μm of HIF-1α inhibitor NS398 in DMEM medium before subjected to the same treatment as group ARC. After the anoxia treatment, each group was reoxygenated in a mixture of 95% air and 5% CO2 incubator for 6 h. Cytoprotections were evaluated by cell viabilities from Trypan blue, lactate dehydrogenase (LDH) release rates, and intracellular cell adhesion molecule-1 (ICAM-1) expressions. Expressions of HIF-1α mRNA and HIF-1α protein from each group were determined by the RT-PCR method and Western blotting, respectively.RESULTS: Ischemia preconditioning increased cell viability, and reduced LDH release and ICAM-1 expressions. Ischemia preconditioning also upregulated the HIF-1α mRNA level and HIF-1α protein expression. However, all of these changes were reversed by HIF-1α inhibitor NS398.CONCLUSION: Ischemia preconditioning effectively inhibited cold hypoxia/warm reoxygenation injury to endothelial cells, and the authors showed for the first time HIF-1α is causally linked to the protective effects of ischemic preconditioning on endothelial cells.