Fortenberry J D, Huber A R, Owens M L
Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
Crit Care Med. 1997 Feb;25(2):303-8. doi: 10.1097/00003246-199702000-00019.
Leukocyte-endothelial cell interactions play a critical role in sepsis-induced multiple organ system failure and acute respiratory distress syndrome. Increased cyclic adenosine 3',5'-monophosphate (cAMP) has been previously reported to inhibit expression of the cytokine-stimulated endothelial cell adhesion molecules, E-selectin, and vascular cell adhesion molecule-1 (VCAM-1). We hypothesized that clinically relevant concentrations of inotropes, such as amrinone and dopamine, which increase cAMP, could inhibit cytokine-stimulated upregulation of endothelial adhesion proteins.
Prospective, controlled in vitro study.
Leukocyte biology laboratory.
Human umbilical vein endothelial cells isolated from neonatal umbilical cord specimens and whole blood obtained from normal human adult volunteers were used in this study.
Endothelial cell monolayers were pretreated with increasing concentrations of amrinone or dopamine, or left untreated as controls, followed by exposure to recombinant human interleukin (IL)-1beta for 6 hrs. Monolayers were then incubated with monoclonal antibodies to E-selectin, VCAM-1, and intercellular adhesion molecule-1 (ICAM-1), fluorescence labeled, and assessed for mean fluorescence intensity by flow cytometry as a measure of surface adhesion molecule concentrations. Whole blood neutrophils were pretreated with or without inotropes, then stimulated with n-formyl methyl leucine phenylalanine. Stimulated neutrophils were incubated with antibodies against the neutrophil adherence protein CD11b and assessed by flow cytometry.
IL-1beta markedly increased E-selectin (p = .01), VCAM-1 (p < .01), and ICAM-1 (p < .001) concentrations (n = 6). Pretreatment with amrinone significantly decreased endothelial E-selectin surface values at all concentrations (p < .001 by analysis of variance, n = 5), including therapeutic concentration ranges. Amrinone also inhibited upregulation of ICAM-1 (p < .001) at therapeutic concentrations, and VCAM-1 (p < .001) at higher concentrations. Dopamine inhibited only E-selectin at relevant concentrations. Neutrophil pretreatment with inotropes did not prevent CD11b upregulation.
Pretreatment with amrinone, and to a lesser degree, with dopamine, at clinically relevant concentrations inhibits in vitro IL-1alpha-induced increases in human umbilical vein endothelial cell adhesion molecule concentrations. Future studies are necessary to investigate the mechanisms of these effects and to determine in vivo efficacy of inotropes as anti-inflammatory agents.
白细胞与内皮细胞的相互作用在脓毒症诱导的多器官系统衰竭和急性呼吸窘迫综合征中起关键作用。先前有报道称，环磷酸腺苷（cAMP）水平升高可抑制细胞因子刺激的内皮细胞黏附分子、E选择素和血管细胞黏附分子-1（VCAM-1）的表达。我们推测，临床上使用的如氨力农和多巴胺等具有正性肌力作用的药物，可升高cAMP水平，从而抑制细胞因子刺激引起的内皮黏附蛋白上调。
前瞻性对照体外研究。
白细胞生物学实验室。
本研究使用从新生儿脐带标本中分离的人脐静脉内皮细胞和从正常成人志愿者获取的全血。
用浓度递增的氨力农或多巴胺预处理内皮细胞单层，或不做处理作为对照，随后暴露于重组人白细胞介素（IL）-1β 6小时。然后将单层细胞与抗E选择素、VCAM-1和细胞间黏附分子-1（ICAM-1）的单克隆抗体孵育，进行荧光标记，并通过流式细胞术评估平均荧光强度，以此作为表面黏附分子浓度的指标。全血中性粒细胞在有或无正性肌力药物预处理的情况下，用N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸刺激。将受刺激的中性粒细胞与抗中性粒细胞黏附蛋白CD11b的抗体孵育，并通过流式细胞术进行评估。
IL-1β显著增加E选择素（p = 0.01）、VCAM-1（p < 0.01）和ICAM-1（p < 0.001）的浓度（n = 6）。氨力农预处理在所有浓度下均显著降低内皮E选择素的表面值（方差分析，p < 0.001，n = 5），包括治疗浓度范围。氨力农在治疗浓度下还抑制ICAM-1的上调（p < 0.001），在较高浓度下抑制VCAM-1的上调（p < 0.001）。多巴胺在相关浓度下仅抑制E选择素。用正性肌力药物预处理中性粒细胞并不能阻止CD11b的上调。
在临床相关浓度下，氨力农预处理，多巴胺预处理的程度较轻，可在体外抑制IL-1α诱导的人脐静脉内皮细胞黏附分子浓度增加。有必要开展进一步研究以探究这些作用的机制，并确定正性肌力药物作为抗炎药物的体内疗效。
