West Michael A, Koons Ann, Crandall Marie, Skinner Ruby, Worley Marcie, Shapiro Michael B
Department of Surgery, Feinberg School of Medicine, Northwestern University, IL 60611, USA.
J Trauma. 2007 Apr;62(4):805-11. doi: 10.1097/01.ta.0000259267.74789.9b.
We sought to determine whether leukocytes from intensive care unit (ICU) patients have altered ERK and p38 kinase activation and specifically if septic patients manifest changes of endotoxin (lipopolysaccharide [LPS]) tolerance. In vitro pretreatment of monocytes (Mono) with LPS induces LPS tolerance with impaired cytokine release and inhibition of ERK and p38 activation after LPS rechallenge.
We hypothesized that macrophage dysregulation, similar to that seen with in vitro LPS tolerance, occurs in critically ill patients with severe sepsis.
Heparinized whole blood from 16 surgical ICU patients and 16 healthy controls was incubated for 15 minutes +/- 10 ng/mL LPS at 37 degrees C. Mono and neutrophil (polymorphonuclear leukocytes [PMN]) diphospho (active) ERK and p38 kinase activation were determined using flow cytometry with monoclonal antibodies. Results are expressed as mean +/- SEM of basal and percentage change (delta %) in positive cells (delta = LPS stimulated - basal). Chi2 test was used for statistics.
Basal ERK was seen in Mono from all groups, but delta % positive only increased in healthy subjects and systemic inflammatory response syndrome (SIRS) patients. No basal Mono or PMN p38 was seen in healthy controls, but LPS significantly activated p38 in both cell types. Mono from patients with sepsis, but not SIRS, had impaired ERK activation. Both PMN and Mono from patients with SIRS had low basal but high LPS-stimulated p38, whereas p38 activation was impaired in patients with sepsis.
Alterations in mitogen activated protein kinases (MAPK) activation are seen in ICU patients. Leukocytes of septic patients, but not those with SIRS, showed characteristics of LPS tolerance. Assessment of leukocyte MAPK activation may identify and differentiate patients with sepsis from those with SIRS.
我们试图确定重症监护病房（ICU）患者的白细胞是否存在细胞外信号调节激酶（ERK）和p38激酶激活的改变，特别是脓毒症患者是否表现出内毒素（脂多糖[LPS]）耐受性的变化。单核细胞（Mono）在体外经LPS预处理后可诱导LPS耐受性，再次接触LPS后细胞因子释放受损，ERK和p38激活受到抑制。
我们假设在患有严重脓毒症的重症患者中，会出现类似于体外LPS耐受性所见的巨噬细胞失调。
将来自16名外科ICU患者和16名健康对照者的肝素化全血在37℃下与10 ng/mL LPS孵育15分钟（±）。使用单克隆抗体通过流式细胞术测定Mono和中性粒细胞（多形核白细胞[PMN]）的双磷酸化（活性）ERK和p38激酶激活情况。结果以基础值的平均值±标准误以及阳性细胞的百分比变化（δ%）（δ = LPS刺激值 - 基础值）表示。采用卡方检验进行统计学分析。
所有组的Mono中均可见基础ERK，但仅健康受试者和全身炎症反应综合征（SIRS）患者的δ%阳性率增加。健康对照者中未见基础Mono或PMN p38，但LPS可显著激活这两种细胞类型中的p38。脓毒症患者而非SIRS患者的Mono中ERK激活受损。SIRS患者的PMN和Mono基础值均较低，但LPS刺激后的p38较高，而脓毒症患者的p38激活受损。
在ICU患者中可见丝裂原活化蛋白激酶（MAPK）激活的改变。脓毒症患者的白细胞而非SIRS患者的白细胞表现出LPS耐受性特征。评估白细胞MAPK激活情况可能有助于鉴别脓毒症患者与SIRS患者。
