Kriebel Fabian, Wittemann Silke, Hsu Hsin-Yun, Joos Thomas, Weiss Manfred, Schneider E Marion
Department of Experimental Anesthesiology, University Clinic Ulm, Steinhoevelstrasse 9, 89075 Ulm, Germany.
Ann N Y Acad Sci. 2006 Dec;1090:168-76. doi: 10.1196/annals.1378.018.
The current comparative investigation analyses markers of inflammation and apoptosis in peripheral blood of intensive care unit (ICU) patients with postoperative/posttraumatic SIRS (systemic inflammatory response syndrome), sepsis, severe sepsis, or septic shock. Inflammatory markers (C-reactive protein [CRP], cytokines, metalloproteinases [MMPs]) and soluble FAS-Ligand (sCD178) were determined in plasma, and apoptosis-relevant antigens such as active caspase-3, Bcl-2, and sCD178 were quantified in whole-blood cell lysates. These parameters were analyzed daily in 20 postoperative/posttraumatic patients: 2 patients had SIRS, 5 suffered from sepsis (2 died), and 13 had septic shock (5 died). Active caspase-3, Bcl-2, and sCD178 were determined by ELISA and by fluorescence-activated cell sorting (FACS)-array kits using bead-assisted flow cytometry. Cytokines and MMPs were quantified by Luminex-assisted Beadlyte assays. Active caspase-3 was identified in defined samples of whole-blood lysates covering, for example, 5/7, 8/18, and 6/11 consecutive days during the patients' stay on the ICU. Also, sCD178 was detected on successive days. Peaks of active caspase-3 antigen contents in whole blood occurred independently of CRP and inflammatory cytokines such as tumor necrosis factor (TNF)-alpha and IL-6. In addition, high MMPs 1-3, 7-10, and 13 concentrations were detected. Interestingly, active caspase-3 and cell-associated sCD178 were either elevated simultaneously or in a close time window. The same was true for Bcl-2. In conclusion, activation of apoptosis can be determined in whole blood of postoperative/posttraumatic patients by active caspase-3 and by Bcl-2. Pro- and antiapoptotic effects during sepsis may occur independently of peaks in inflammatory markers. Apoptosis could explain modeling and remodeling of leukocyte subpopulations.
当前的比较研究分析了术后/创伤后全身炎症反应综合征（SIRS）、脓毒症、严重脓毒症或感染性休克的重症监护病房（ICU）患者外周血中的炎症和凋亡标志物。在血浆中测定炎症标志物（C反应蛋白[CRP]、细胞因子、金属蛋白酶[MMPs]）和可溶性FAS配体（sCD178），并在全血细胞裂解物中对与凋亡相关的抗原如活性半胱天冬酶-3、Bcl-2和sCD178进行定量。每天对20例术后/创伤后患者的这些参数进行分析：2例患者患有SIRS，5例患有脓毒症（2例死亡），13例患有感染性休克（5例死亡）。活性半胱天冬酶-3、Bcl-2和sCD178通过酶联免疫吸附测定（ELISA）以及使用微珠辅助流式细胞术的荧光激活细胞分选（FACS）阵列试剂盒进行测定。细胞因子和MMPs通过Luminex辅助的Beadlyte检测进行定量。在患者入住ICU期间，在全血细胞裂解物的特定样本中鉴定出活性半胱天冬酶-3，例如连续5/7、8/18和6/11天。此外，连续几天检测到sCD178。全血中活性半胱天冬酶-3抗原含量的峰值独立于CRP以及肿瘤坏死因子（TNF）-α和白细胞介素-6等炎症细胞因子出现。此外，检测到MMPs 1 - 3、7 - 10和13的高浓度。有趣的是，活性半胱天冬酶-3和细胞相关的sCD178要么同时升高，要么在接近的时间窗口内升高。Bcl-2也是如此。总之，通过活性半胱天冬酶-3和Bcl-2可以在术后/创伤后患者的全血中确定凋亡的激活。脓毒症期间的促凋亡和抗凋亡作用可能独立于炎症标志物的峰值而发生。凋亡可以解释白细胞亚群的塑造和重塑。
