Coimbra Raul, Loomis William, Melbostad Heidi, Tobar Maria, Porcides Rafael D, Lall Rohan, Holbrook Troy, Hoyt David B
Division of Trauma, Department of Surgery, University of California San Diego School of Medicine, CA 92103-8896, USA.
J Trauma. 2005 Aug;59(2):257-64; discussion 264-5. doi: 10.1097/01.ta.0000174678.12523.9f.
Hypertonic saline (HS) and pentoxifylline (PTX) have been shown to modulate polymorphonuclear neutrophil (PMN) functions after shock and sepsis. We hypothesized that a combination of HS and PTX (HSPTX) would down-regulate PMN functions and inflammatory mediator synthesis more effectively than each alone, possibly by acting at different steps of the signaling pathways, ultimately leading to an enhanced effect.
Whole blood from healthy volunteers was stimulated with lipopolysaccharide (LPS) (100 microg/mL), f-methionyl-leucyl-phenylalanine (1 micromol/L), and phorbol 12-myristate 13-acetate (1 microg/mL). Baseline oxidative burst was measured by flow cytometry. Two different concentrations of NaCl to achieve increases of 10 mmol/L (HS10) and 40 mmol/L (HS40) above isotonicity, simulating increases in sodium levels seen after infusion of 3% HS and 7.5% HS, were used. PTX (2 mmol/L), HS10, HS40, HSPTX10, and HSPTX40 were added to whole blood concomitantly to the activators. PMN CD14 and CD11b expression were measured by flow cytometry and tumor necrosis factor-alpha levels by enzyme-linked immunosorbent assay in LPS-stimulated whole blood.
The combination of PTX with HS10 and with HS40 markedly decreased LPS- (27 +/- 7 and 23 +/- 6 vs. 100; p < 0.01), f-methionyl-leucyl-phenylalanine- (54 +/- 11 and 55 +/- 8 vs. 100; p < 0.05), and phorbol 12-myristate 13-acetate- (30 +/- 4 and 54 +/- 9 vs. 100; p < 0.01 and p < 0.05, respectively) induced PMN oxidative burst. Furthermore, a significant decrease in LPS-induced neutrophil CD11b expression after PTX treatment (79 +/- 5 vs. 100; p < 0.05) and HSPTX40 (68 +/- 7 vs. 100; p < 0.05) was observed. HSPTX10 (7 +/- 0.6; p < 0.001) or HSPTX40 (6 +/- 1.4; p < 0.001) markedly decreased tumor necrosis factor-alpha production to levels similar to those observed with PTX alone (6.5 +/- 0.5; p < 0.001) and significantly lower than HS10 (110 +/- 4.9; p < 0.001) and HS40 alone (83 +/- 1.5; p < 0.001)
HSPTX down-regulates neutrophil activation and proinflammatory mediator synthesis more effectively that HS alone. HSPTX may have significant applications as a novel fluid resuscitation strategy in hemorrhagic shock.
高渗盐水（HS）和己酮可可碱（PTX）已被证明可在休克和脓毒症后调节多形核中性粒细胞（PMN）功能。我们假设HS和PTX联合使用（HSPTX）比单独使用更能有效下调PMN功能和炎症介质合成，可能是通过作用于信号通路的不同步骤，最终产生增强效应。
用脂多糖（LPS）（100微克/毫升）、f-甲硫氨酰-亮氨酰-苯丙氨酸（1微摩尔/升）和佛波醇12-肉豆蔻酸酯13-乙酸酯（1微克/毫升）刺激健康志愿者的全血。通过流式细胞术测量基线氧化爆发。使用两种不同浓度的氯化钠，使渗透压分别比等渗增加10毫摩尔/升（HS10）和40毫摩尔/升（HS40），模拟输注3%HS和7.5%HS后钠水平的升高。将PTX（2毫摩尔/升）、HS10、HS40、HSPTX10和HSPTX40与激活剂同时加入全血中。通过流式细胞术测量PMN的CD14和CD11b表达，并通过酶联免疫吸附测定法测量LPS刺激的全血中肿瘤坏死因子-α水平。
PTX与HS10和HS40联合使用显著降低了LPS（分别为27±7和23±6 vs. 100；p<0.01）、f-甲硫氨酰-亮氨酰-苯丙氨酸（分别为54±11和55±8 vs. 100；p<0.05）以及佛波醇12-肉豆蔻酸酯13-乙酸酯（分别为30±4和54±9 vs. 100；p分别<0.01和p<0.05）诱导的PMN氧化爆发。此外，观察到PTX处理后（79±5 vs. 100；p<0.05）和HSPTX40处理后（68±7 vs. 100；p<0.05）LPS诱导的中性粒细胞CD11b表达显著降低。HSPTX10（7±0.6；p<0.001）或HSPTX40（6±1.4；p<0.001）显著降低肿瘤坏死因子-α的产生，降至与单独使用PTX时相似的水平（6.5±0.5；p<0.001），且显著低于单独使用HS10（110±4.9；p<0.001）和HS40（83±1.5；p<0.001）。
HSPTX比单独使用HS更能有效下调中性粒细胞活化和促炎介质合成。HSPTX作为一种新型的液体复苏策略在失血性休克中可能有重要应用。
