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Biological response of human aortic endothelial cells exposed to acellular hemoglobin solutions developed as potential blood substitutes.

机译:暴露于脱细胞血红蛋白溶液中的人类主动脉内皮细胞的生物学反应已发展为潜在的血液替代品。

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The cardiovascular effects of hemoglobin-based oxygen carriers (HBOCs) are mainly related to their nitric oxide (NO) scavenging properties but other effects such as the impact of these hemoglobins on the endothelial cell (EC) biology are not well understood. We hypothesized that HBOCs could modify EC functions by altering gene expression, in particular the endothelial NO synthase (NOS3) and/or by activating EC. Cultured human aortic endothelial cells (HAEC) were incubated for 3 hours with purified cell-free Hb, Dex-BTC-Hb or alpha alpha-Hb (16 g/L). Expression of NOS3 mRNA and protein were assessed by semi-quantitative RT-PCR and Western blot respectively immediately after and 24 hours after incubation. The expression and localization of the adhesion molecule ICAM-1 were detected by fluorescence microscopy. None of the solutions tested modified NOS3 mRNA and protein expression despite adequate controls that up- or down-regulate NOS3 expression. The expression and the localization of ICAM-1 on the cell membrane were modified after 3 hours of incubation with all the hemoglobin solutions tested in a manner similar to tumor necrosis factor-alpha. In conclusion, HAEC incubation with clinically relevant concentrations of HBOCs induced changes in the pattern of ICAM-1 expression consistent with cell activation/cell signaling mechanisms. However, HBOCs did not alter NOS3 gene expression.
机译:基于血红蛋白的氧载体(HBOC)的心血管作用主要与其一氧化氮(NO)清除特性有关,但诸如此类血红蛋白对内皮细胞(EC)生物学的影响之类的其他作用尚不清楚。我们假设HBOCs可以通过改变基因表达,尤其是内皮型NO合酶(NOS3)和/或激活EC来修饰EC功能。将培养的人主动脉内皮细胞(HAEC)与纯化的无细胞Hb,Dex-BTC-Hb或alpha alpha-Hb(16 g / L)孵育3小时。孵育后立即和孵育后24小时分别通过半定量RT-PCR和蛋白质印迹法评估NOS3 mRNA和蛋白的表达。通过荧光显微镜检测粘附分子ICAM-1的表达和定位。尽管有适当的控制可以上调或下调NOS3表达,但没有一种测试的溶液能修饰NOS3 mRNA和蛋白质表达。用所有测试的血红蛋白溶液孵育3小时后,以类似于肿瘤坏死因子-α的方式修饰ICAM-1在细胞膜上的表达和定位。总之,HAEC与临床相关浓度的HBOC孵育可诱导ICAM-1表达模式的变化,这与细胞激活/细胞信号传导机制一致。但是,HBOCs不会改变NOS3基因的表达。

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