【摘要】 目的研究缺氧预适应对内皮细胞冷缺氧/温复氧损伤的作用及可能机制。方法培养的内皮细胞ECV-304分成4组,A组,对照组;B组,冷缺氧/温复氧(A/R)组95%N2/5%CO2缺氧装置中用4℃UW液保存24h;C组,缺氧预适应组(APC),内皮细胞在缺氧前遭受4个循环短时间缺氧/复氧;D组,缺氧诱导因子-1α(HIF-1α)抑制组缺氧预适应前,细胞培养液中加入5μmol/L的HIF-1α抑制剂NS398,余同C组。缺氧结束后,各组37℃5%CO295%空气中复氧6h。分别以逆转录-聚合酶链反应(RT-PCR)法、Western blot法检测各组HIF-1αmRNA和蛋白表达,内皮细胞保护作用通过细胞存活率(台盼蓝拒染法)、LDH释出率及ICAM-1的表达判定。结果缺氧预适应明显上调HIF-1αmRNA和蛋白表达,显著抑制冷保存内皮细胞缺氧/复氧损伤,表现为更高的细胞存活率,更少的LDH释出和ICAM-1的表达。而HIF-1α抑制剂可逆转这种保护作用。结论缺氧预适应能有效地抑制冷保存内皮细胞缺氧/复氧损伤,这种细胞保护作用可能是通过HIF-1α介导的。更多还原

【Abstract】 Objective To investigate the effect and mechanism of anoxia-preconditioning on cold anoxia/warm reoxygenation injury to endothelial cells.Methods Cultured endothelial cells were divided into 4 groups:Group A as control;Group B,the cells were preserved in 40 ℃ UW solution and subjected to 24 h-anoxia.Group C,hypoxia-preconditioning was performed by 4 cycles’ short anoxia and reoxygenation before prolonged anoxia.Group D,pretreating the cells with HIF-1α inhibitor,NS398.After anoxia,the cells were exposed to 37 ℃ 5% CO₂ 95% air for 6 h.HIF-1 αmRNA and protein expression was respectively determined by RT-PCR and Western blot.The cytoprotection was evaluated by Dyed rate of Typan blue,LDH release and ICAM-1 expression.Results Anoxia-preconditioning upregulated HIF-1α mRNA and augmented HIF-1α protein.The effect was accompanied by improved cell viability,reduced LDH release and ameliorated ICAM-1 expression.However,the effect was cancelled by NS398.Conclusion Anoxia-preconditioning can provide the protection to endothelial cell from cold hypoxia/warm reoxygenation injury,and the effect may be mediated by HIF-1α.更多还原