Toussaint-Hacquard M, Devaux Y, Longrois D, Faivre-Fiorina B, Muller S, Stoltz J F, Vigneron C, Menu P
Laboratoire d'Hématologie-Physiologie, UPRES EA 3452, Faculté de Pharmacie, 54000 Nancy, France.
Life Sci. 2003 Jan 24;72(10):1143-57. doi: 10.1016/s0024-3205(02)02368-8.
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.
基于血红蛋白的氧载体（HBOCs）的心血管效应主要与其一氧化氮（NO）清除特性有关，但这些血红蛋白对内皮细胞（EC）生物学的其他影响，如对内皮细胞功能的影响尚不清楚。我们推测，HBOCs可能通过改变基因表达，特别是内皮型一氧化氮合酶（NOS3）的表达和/或激活内皮细胞来改变内皮细胞功能。将培养的人主动脉内皮细胞（HAEC）与纯化的无细胞血红蛋白、右旋糖酐-双（三甲基氨基）钴血红蛋白或αα-血红蛋白（16 g/L）孵育3小时。孵育后立即及24小时后，分别通过半定量逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法评估NOS3 mRNA和蛋白质的表达。通过荧光显微镜检测黏附分子细胞间黏附分子-1（ICAM-1）的表达和定位。尽管有上调或下调NOS3表达的适当对照，但所测试的溶液均未改变NOS3 mRNA和蛋白质的表达。与肿瘤坏死因子-α类似，用所有测试的血红蛋白溶液孵育3小时后，细胞膜上ICAM-1的表达和定位发生了改变。总之，用临床相关浓度的HBOCs孵育HAEC会导致ICAM-1表达模式发生变化，这与细胞激活/细胞信号传导机制一致。然而，HBOCs并未改变NOS3基因的表达。
