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The in vivo kinetics of tissue factor messenger RNA expression during human endotoxemia: relationship with activation of coagulation

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BLOOD 2000年第2期96卷 554-559页

作者： Franco, RF de Jonge, E Dekkers, PEP Timmerman, JJ Spek, CA van Deventer, SJH van Deursen, P van Kerkhoff, L van Gemen, B ten Cate, H van der Poll, T Reitsma, PH Univ Amsterdam Acad Med Ctr Lab Expt Internal Med NL-1105 AZ Amsterdam Netherlands Univ Amsterdam Acad Med Ctr Dept Intens Care NL-1105 AZ Amsterdam Netherlands Organon Teknika BV Boxtel Netherlands

Triggering of the tissue factor (TF)-dependent coagulation pathway is considered to underlie the generation of a procoagulant state during endotoxemia, To determine the in vivo pattern of monocytic TF messenger RNA (mRNA) expression during endotoxemia, 10 healthy volunteers were injected with lipopolysaccharide (LPS, 4 ng/kg) and blood was collected before and 0.5, 1, 2, 3, 4, 6, 8, and 24 hours after LPS administration. Total blood RNA was isolated and amplified by NASBA (nucleic acid sequence-based amplification), followed by quantitation of TF mRNA by an electrochemiluminescence (ECL) assay, To com-pare the pattern of coagulation activation with the kinetics of monocytic TF mRNA expression, we measured plasma levels of markers of thrombin generation, thrombin-antithrombin (TAT) complexes, and prothrombin fragment 1 + 2 (F1 + 2). Baseline value (mean +/- SEM) of the number of TF mRNA molecules per monocytic cell was 0.08 +/- 0.02. A progressive and significant (P < .0001) increase in TF expression was observed after LPS Injection (+0.5 hour: 0.3 +/- 0.1, +1 hour: 1.3 +/- 0.9, +2 hours: 4.1 +/- 0.9), peaking at +3 hours (10 +/- 1.9 TF mRNA molecules per monocyte), As TF mRNA levels Increased, thrombin generation was augmented, Peak levels of TAT and F1 + 2 were reached later (at t +4 hours) than those of TF mRNA. TF mRNA, TAT, and F1 + 2 levels returned to baseline after 24 hours. In conclusion, we used a NASBA/ECL-based technique to quantify TF mRNA in whole blood during human endotoxemia and observed a 125-fold increase in TF mRNA levels, Our data demonstrate a pivotal role for enhanced TF gene activity in the activation of coagulation after LPS challenge. (C) 2000 by The American Society of Hematology.

关键词：抗凝血酶Ⅲ/分析血液凝固内毒素血症/血液基因表达动力学脂多糖类/药理学肽碎片/分析肽水解酶类/分析凝血酶原/分析 RNA 信使/代谢凝血致活酶/遗传学凝血致活酶/代谢成年人人类男(雄)性

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Vitamin E in relation to lipid peroxidation in experimental septic shock

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PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS 2000年第3期62卷 195-199页

作者： Basu, S Eriksson, M Uppsala Univ Fac Med Dept Geriatr Clin Nutr Res SE-75125 Uppsala Sweden Uppsala Univ Fac Med Dept Anaesthesiol & Intens Care SE-75125 Uppsala Sweden

Lipid peroxidation and antioxidant balance in the body is a crucial factor in the pathophysiology of various diseases. This study investigates the circulatory alpha-tocopherol levels and its relationship with 8-iso-prostaglandin F(2 alpha) (8-iso-PGF(2 alpha)), a non-enzymatic and, 15-keto-13,14-dihydro-PGF(2 alpha) (15-K-DH-PGF(2 alpha)), a cyclooxygenase catalysed oxidation product of arachidonic acid in experimental septic shock in pigs. A steady decrease in alpha-tocopherol levels in plasma was observed in both survivor and non-survivor animals. A simultaneous increase of oxidative injury indicator, plasma 8-iso-PGF(2 alpha) was seen in both groups but with a different fashion. 8-Iso-PGF(2 alpha) levels increased steadily in the animals that died during the experiment. An early and rapid increase of plasma 15-K-DH-PGF(2 alpha), an inflammatory response indicator, was also observed in all animals. There was a significant difference in the kinetics of decrement of cx-tocopherol levels and a concomitant increase in 15-K-DH-PGF(2 alpha) levels among the non-survivors. Thus, a successive disappearance of circulatory vitamin E in conjunction with the surge of plasma isoprostanes and prostaglandins impairs the oxidant-antioxidant balance in favour of the former and may possibly have an effect on the survivality during experimental porcine septicaemia. (C) 2000 Harcourt Publishers Ltd.

关键词：花生四烯酸/血液地诺前列素/类似物和衍生物地诺前列素/血液内毒素血症/血液内毒素血症/酶学内毒素血症/病理生理学类地诺前列素脂质过氧化作用氧化性应激前列腺素内过氧化物合酶类/血液休克脓毒性/血液休克脓毒性/酶学休克脓毒性/病理生理学猪血管收缩药/血液维生素E/血液维生素E/生理学动物女(雌)性男(雄)性

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Tissue factor pathway inhibitor dose-dependently inhibits coagulation activation without influencing the fibrinolytic and cytokine response during human endotoxemia

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BLOOD 2000年第4期95卷 1124-1129页

作者： de Jonge, E Dekkers, PEP Creasey, AA Hack, CE Paulson, SK Karim, A Kesecioglu, J Levi, M van Deventer, SJH van der Poll, T Univ Amsterdam Dept Intens Care Acad Med CtrLab Expt Internal Med Cent LabRed Cross Blood Transfus Serv NL-1100 DD Amsterdam Netherlands Univ Amsterdam Dept Vasc Med Acad Med Ctr Cent LabRed Cross Blood Transfus Serv NL-1100 DD Amsterdam Netherlands

Inhibition of the tissue factor pathway has been shown to attenuate the activation of coagulation and to prevent death In a gram-negative bacteremia primate model of sepsis, It has been suggested that tissue factor influences inflammatory cascades other than the coagulation system. The authors sought to determine the effects of 2 different doses of recombinant tissue factor pathway inhibitor (TFPI) on endotoxin-induced coagulant, fibrinolytic, and cytokine responses in healthy humans. Two groups, each consisting of 8 healthy men, were studied in a double-blind, randomized, placebo-controlled crossover study. Subjects were studied on 2 different occasions. They received a bolus intravenous injection of 4 ng/kg endotoxin, which was followed by a 6-hour continuous infusion of TFPI or placebo. Eight subjects received 0.05 mg/kg per hour TFPI after a bolus of 0.0125 mg/kg (tow-dose group), and 8 subjects received 0.2 mg/kg per hour after a bolus of 0.05 mg/kg (high-dose group). Endotoxin injection induced the activation of coagulation, the activation and subsequent inhibition of fibrinolysis, and the release of proinflammatory and antiinflammatory cytokines. TFPI infusion induced a dose-dependent attenuation of thrombin generation, as measured by plasma F1 + 2 and thrombin-antithrombin complexes, with a complete blockade of coagulation activation after high-dose TFPI. Endotoxin-induced changes in the fibrinolytic system and cytokine levels were not altered by either low-dose or high-dose TFPI, The authors concluded that TFPI effectively and dose-dependently attenuates the endotoxin-induced coagulation activation in humans without influencing the fibrinolytic and cytokine response.

关键词：抗凝药/投药和剂量抗凝药/药理学抗凝血酶Ⅲ/代谢血液凝固/药物作用血压/药物作用体温/药物作用交叉研究细胞因子类/血液双盲法用药计划表内毒素血症/血液内毒素血症/药物疗法内毒素血症/免疫学大肠杆菌感染/血液大肠杆菌感染/药物疗法大肠杆菌感染/免疫学纤维蛋白溶解/药物作用心率/药物作用输注静脉内注射静脉内脂多糖类/投药和剂量脂蛋白类/投药和剂量脂蛋白类/药理学肽水解酶类/代谢纤溶酶原激活物抑制物1/血液成年人人类男(雄)性

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Concentrations of lipopolysaccharide-binding protein, bactericidal/permeability-increasing protein, soluble CD14 and plasma lipids in relation to endotoxaemia in patients with alcoholic liver disease

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ALCOHOL AND ALCOHOLISM 2002年第1期37卷 81-86页

作者： Schäfer, C Parlesak, A Schütt, C Bode, JC Bode, C Robert Bosch Krankenhaus Dept Internal Med Div Gastroenterol D-70376 Stuttgart Germany Univ Hohenheim Dept Physiol Nutr D-70599 Stuttgart Germany Ernst Moritz Arndt Univ Greifswald Dept Immunol D-17489 Greifswald Germany

There is increasing evidence that gut leakage in persons with chronic alcohol misuse leads to endotoxaemia, which might contribute to the development of alcoholic hepatitis or cirrhosis. In addition, it was recently shown that the endotoxin-binding capacity of whole blood is reduced in these patients. To analyse this phenomenon, we measured the concentration of functionally important endotoxin-binding plasma components which modify the action of endotoxin. In patients with minimal (n = 10), intermediate (n = 9), and cirrhotic alcoholic liver disease (n = 11), and healthy controls (n = 11), plasma endotoxin was determined in a limulus assay. The concentration of lipoproteins was assessed by measuring apolipoproteins, the other factors were directly measured in immunoassays. In the entire group of alcoholics, endotoxin and the concentration of binding factors that are involved in the action of endotoxin on its target cells (LPS-binding protein and sCD14) were increased. Endotoxin antagonists, such as bactericidal/permeability-increasing protein and high-density lipoprotein, were increased in the pre-cirrhotic stages, whereas a significant reduction of the latter was observed in cirrhosis. Low-density lipoprotein remained unchanged. The elevation of binding factors in the pre-cirrhotic stages of alcoholic liver disease might attenuate the effects of endotoxaemia, whereas in cirrhosis the reduction of high density lipoprotein, to which large quantities of endotoxin bind, may contribute to its pro-inflammatory effects.

关键词：急相蛋白质类慢性酒精中毒/血液慢性酒精中毒/病因学抗微生物阳离子肽类血蛋白质类/分析载体蛋白质类/血液色谱法内毒素血症/血液内毒素血症/并发症酶联免疫吸附测定免疫测定脂类/血液肝疾病酒精性/血液肝疾病酒精性/并发症膜糖蛋白类膜蛋白质类成年人女(雌)性人类男(雄)性中年人

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Peroxisome proliferator-activated receptor γ1 expression in porcine white blood cells:: Dynamic regulation with acute endotoxemia

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 1999年第3期263卷 749-753页

作者： Leininger, MT Portocarrero, CP Houseknecht, KL Purdue Univ Dept Anim Sci W Lafayette IN 47907 USA

Peroxisome proliferator-activated receptor gamma (PPAR gamma), a primary regulator of adipocyte differentiation, has been implicated in the regulation of monocyte and macrophage function in vitro. We report that PPAR gamma protein is expressed in porcine peripheral white blood cells (WBC), and that PPAR gamma 1 but not gamma 2 mRNA predominates. Additionally, we provide the first evidence that in vivo lipopolysaccharide challenge (LPS, 25 mu g/kg BW) causes a dynamic increase in PPAR gamma protein expression in peripheral WBC (P < 0.05). PPAR gamma expression was increased 2-fold over basal (1 hr post-LPS), was maximal by 4 hr (3-fold), and was normalized to control by 8 hr post-LPS. Changes in PPAR gamma expression coincided with or closely followed LPS-induced changes in plasma cortisol, TNF-alpha, insulin, IGF-1, glucose, and free fatty acids. These data suggest that induction of PPAR gamma expression in WBC may play a role in host response to acute inflammatory challenge and may prove to be an important target of anti-inflammatory therapies. (C) 1999 Academic Press.

关键词： DNA结合蛋白质类/血液 DNA结合蛋白质类/遗传学内毒素血症/血液基因表达调控/药物作用基因表达调控/生理学动力学白细胞/药物作用白细胞/代谢脂多糖类/毒性睾丸切除术蛋白质亚型/血液蛋白质亚型/遗传学 RNA 信使/遗传学受体胞质和核/血液受体胞质和核/遗传学参考值猪时间因素转录因子/血液转录因子/遗传学转录遗传/药物作用动物男(雄)性

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