Antithrombin reduces mesenteric venular leukocyte interactions and small intestine injury in endotoxemic rats

We examined the hypothesis that recombinant human antithrombin would reduce mesenteric venule leukocyte adhesion and small intestine injury in endotoxemic rats. Endotoxemic (endotoxin 10 mg/kg, intravenously) rats were treated either with saline or recombinant human antithrombin (250 and 500 U/kg). In some rats, indomethacin (100 mg/kg, intraperitoneally) was injected 60 min prior to endotoxin and recominant human antithrombin (500 U/kg) treatment. Compared to controls, intravital videomicroscopy of the mesentric venule showed an increase of leukocyte rolling (55+/-17 versus 70+/-19 leukocytes/min; P < 0.05) and firm adhesion (1.1+/-0.3 versus 5.8+/-0.8 leukocytes/100 microm; P < 0.05) in endotoxemic rats. Recombinant human antithrombin attenuated endotoxin-induced venular endothelium leukocyte adhesive cascade. The beneficial effects of recombinant human antithrombin on leukocyte adhesion were inhibited by indomethacin (100 mg/kg, intraperitoneally) in endotoxemic rats. Endotoxin treatment increased fluorescein isothiocyanate (FITC)-labeled dextran 4,000 (FD4) gut lumen to plasma ratio and wet weight/dry weight ratio. Recombinant human antithrombin (500 U/kg) attenuated endotoxin-induced gut injury. These observations suggest that recombinant human antithrombin reduces endothelium-leukocyte interactions in endotoxemic rats by interacting with local prostacyclin production.     

Inhibition of glucocorticoid receptor binding by nitric oxide in endotoxemic rats

OBJECTIVE: Nitric oxide is an important participant in septic shock. For example, it causes profound vasodilation and hypotension. Despite their potent antiinflammatory properties, glucocorticoids are not routinely used in septic shock. Some studies show that antiinflammatory doses of glucocorticoids can be beneficial, but other studies do not indicate their use in this situation. We have previously shown the inhibitory effect of nitric oxide on glucocorticoid receptor binding in vitro. Nitric oxide donors decreased the binding of immunoprecipitated glucocorticoid receptor obtained from mouse L929 fibroblasts. These in vitro findings prompted us to study whether in vivo manipulations of the nitric oxide system would interfere with the glucocorticoid receptor binding. DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university. SUBJECTS: Female Wistar rats. INTERVENTIONS: Injection of bacterial lipopolysaccharide, anesthesia, cardiovascular perfusion, and organ removal for biochemical assays. MEASUREMENTS AND MAIN RESULTS: Following lipopolysaccharide injection, plasma nitrate + nitrite increased and inducible nitric oxide synthase activity was stimulated in several organs, the highest rates being in the lung and spleen. If dexamethasone was injected before lipopolysaccharide, it completely blocked inducible nitric oxide synthase induction and the increase in plasma nitrate + nitrite. On the other hand, if dexamethasone was injected after lipopolysaccharide, it failed to affect both inducible nitric oxide synthase induction and increased plasma nitrate + nitrite levels. Lipopolysaccharide also caused an inhibition of glucocorticoid receptor binding in lung and spleen. Previous administration of a nitric oxide synthase inhibitor prevented both lipopolysaccharide-induced decrease in glucocorticoid receptor binding and the increase in plasma nitrate + nitrite. Injection of a nitric oxide donor into naive animals significantly decreased glucocorticoid receptor binding activity and prevented dexamethasone-induced increase in liver tyrosine aminotransferase activity. CONCLUSIONS: The results indicate that the failure of glucocorticoids to exhibit their antiinflammatory effects when administered to endotoxemic rats may be explained, at least in part, by the nitric oxide-induced inhibition of glucocorticoid receptor binding ability, thus precluding the expression of the antiinflammatory effects of both exogenous and endogenous corticosteroids.     

Inhaled nitric oxide in ARDS

The role of nitric oxide (NO) in numerous physiologic systems only recently has been discovered. When used as a gas, inhaled NO (iNO) has many unique properties that cause immediate improvements in pulmonary hemodynamics and oxygenation. Acute benefits in physiologic parameters have been demonstrated in numerous studies of iNO in acute respiratory distress syndrome (ARDS), but recent randomized controlled trials have failed to show improvement in outcome. The addition of other treatments that prolong or enhance the affect of iNO or its use with other ventilator modalities such as prone positioning or high-frequency ventilation offer hope that iNO may be beneficial in select groups of patients.     

Inhaled nitric oxide therapy]

Inhaled nitric oxide (INO) therapy is becoming an indispensable measure in some critically ill patients with pulmonary hypertension. Most importantly, it has been shown that INO significantly reduces the necessity for extracorporeal lung assist in newborns with hypoxemic respiratory failure and pulmonary hypertension. A large number of basic and clinical studies are making great steps in delineating its physiology, techniques, side effects, and clinical efficacy. This article reviews the current knowledge of INO therapy including a novel device of nitric oxide production system.     

Intestinal nitric oxide in the normal and endotoxemic pig

The gut is considered a central organ in the pathogenesis of sepsis and multiple organ failure, where several mediators, including endothelin (ET) and nitric oxide (NO), are involved. The aim of the current study was to characterize, by direct measurements, the intestinal NO production in the anesthetized pig during normal and endotoxemic conditions. In pigs subjected to endotoxin infusion, there was a progressive decrease in jejunal luminal NO levels, as well as portal venous blood flow and blood pressure. The ET- blocker 4-tert-butyl-N-[6-(2-hydroxy-ethoxy)-5-(2-methoxy-phenoxy)-2,2'-bipyrimidin-4-yl]-benzenesulfonamide (bosentan) completely reversed the reduction in portal venous blood flow without affecting intestinal NO levels. In control pigs, the NO synthase inhibitor N(omega)-nitro-L-arginine methyl ester dose-dependently decreased intestinal NO levels and mesenteric blood flow--effects that were reversed by L-arginine. We conclude that intestinal NO is a product of mucosal NO synthase activity, and is profoundly decreased during endotoxemia in the pig.     

Prolactin secretion in hypothyroid endotoxemic rats: involvement of L-arginine and nitric oxide synthase

The identification of nitric oxide (NO) within the hypothalamus and pituitary gland has suggested its role as modulator of the activity on hypothalamic-pituitary axis. Hypothalamic NO synthase (NOS) is known to be regulated by thyroid hormones. We investigated the effects of previous injection of N-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, and L-arginine (L-Arg), the substrate for NO synthesis, on prolactin (PRL) secretion, through the lipopolysaccharide (LPS)-induced inflammatory response in thyroidectomized (TX) rats. TX or sham-operated (N) rats were intraperitoneally (i.p.) injected with L-NAME (10 mg kg) or L-Arg (200 mg kg) or the same volume of vehicle (saline solution) 30 min before endotoxemia-induction with LPS at 250 mug (100 g body weight), i.p.. In N rats, NO increased PRL release in response to endotoxemia, whereas in hypothyroid rats, NO appeared to have the opposite effect. Our data support the hypothesis that NO exerts a modulatory influence on PRL secretion after LPS-induced inflammatory response.     

Inhaled nitric oxide in infants and children

NO has been used successfully to treat PPHN, reducing the need for ECMO. NO has also been used in the cardiac catheterization laboratory to determine if pulmonary hypertension will decrease with NO. Patients who do not respond to NO are at higher risk after open-heart surgery, because their pulmonary hypertension will be difficult to treat. Postoperatively, NO can be used to determine if pulmonary hypertension is caused by vasoconstriction or by an obstruction. Inhaled Nitric Oxide at a Glance: ACTION: Selective pulmonary vasodilation without systemic vasodilation. USE: Treatment of pulmonary hypertension. CONCENTRATION AND ROUTE: Lowest concentration that will produce pulmonary vasodilation and improved oxygenation. Concentration should be kept < 80 ppm. CONTRAINDICATION: Neonate that is ductal-dependent. TOXIC EFFECTS: Keep methemoglobin level < 5%. Keep nitric dioxide, which can cause lung damage, < 7 ppm. Risk of bleeding. MONITOR: Levels of NO/NO2. Platelets. Arterial blood gas (ABG). Methemoglobin. WEANING: Decrease NO by 20%, monitoring ABG at 3- to 4-hour intervals. If there is a decrease in oxygenation, increase NO. Increase FIO2 20% when NO is discontinued. Unsuccessful treatment with NO--keep on NO until ECMO is available.     

Inhaled nitric oxide delivery and monitoring

Inhaled nitric oxide (NO) was found to cause selective pulmonary vasodilation in the late 1980's and since then there has been a huge interest in studying its clinical benefits. The equipment used to deliver and monitor inhaled NO has gone through a dramatic evolution from simple flow meters and industrial monitors to to-days purpose built, fully integrated, NO delivery and monitoring systems that were designed specifically for the demanding area of the intensive care unit. This paper explores the evolution of inhaled NO delivery systems and identifies the design challenges, the safety and regulatory requirements and the ease of use issues that had to be solved to bring this new exciting new class of medical device in to clinical use.     

Inhaled nitric oxide down-regulates intrapulmonary nitric oxide production in lipopolysaccharide-induced acute lung injury

OBJECTIVE: To examine whether inhaled nitric oxide (NO) affected the intrapulmonary production of NO, reactive oxygen species, and nuclear factor-kappaB in a lipopolysaccharide (LPS)-induced model of acute lung injury. DESIGN: Prospective, randomized, laboratory study. SETTING: Experimental laboratory at a biomedical institute. SUBJECTS: Twenty male rabbits weighing 2.5-3.5 kg. INTERVENTIONS: Saline or LPS (5 mg/kg of body weight) was administered intravenously with or without NO inhalation (10 ppm) in each group of five rabbits. MEASUREMENTS AND MAIN RESULTS: LPS increased the lung leak index, the neutrophils and NO levels in bronchoalveolar lavage fluid, and NO levels produced by resting and stimulated alveolar macrophages. Inhaled NO decreased the lung leak index, the neutrophils and NO levels as measured by nitrite levels in the lavage fluid, and NO produced by the resting and stimulated alveolar macrophages. Inhaled NO also blocked the activities of reactive oxygen species and nuclear factor-kappaB binding to DNA in lavage cells and in alveolar macrophages. CONCLUSION: Inhaled NO attenuates LPS-induced acute lung injury, possibly by decreasing NO production in the lungs. The mechanism of reducing NO production resulting from inhaled NO may involve, in part, the activities of reactive oxygen species and/or nuclear factor-kappaB.     

外源性一氧化氮吸入治疗急性呼吸窘迫综合征

目的　探讨对于急性呼吸窘迫综合征 (ARDS)病人采用吸入外源性一氧化氮 (NO)治疗引起血流动力学和呼吸功能的改变。方法　 8例ARDS患者行机械通气、漂浮导管 ,吸入NO 2 0ppm ,观察 3个时相点 :①基础水平 ;②开始吸入NO后 2 0min ;③停止吸入NO后 2 0min ;④血流动力学、血气和肺内分流量的变化。结果　吸入NO后 ,患者平均肺动脉压 (MPAP)从 (3 74± 0 6 4 )kPa下降至 (2 6 1± 0 5 1)kPa(P <0 0 1) ,而平均动脉压 (MAP)、肺毛细血管楔压及中心静脉压无明显变化 ;同时 ,动脉血氧分压、动脉血氧饱和度分别从 (10 3± 1 81)kPa和 (93 4± 3 3) %上升至 (11 8± 1 0 7)kPa和 (96 0± 2 3) % (P <0 0 5 ,P <0 0 1) ,而肺内分流量从 (30 8± 7 4 ) %下降至 (2 3 5± 3 5 ) % (P <0 0 1) ;停止吸入NO 2 0min后 ,各指标又基本恢复至基础水平。结论　吸入外源性NO具有较好的肺血管扩张选择性 ,在降低肺动脉压的同时能改善气体交换功能     

Potentiation of anandamide effects in mesenteric beds isolated from endotoxemic rats

The aim of the present experiments was to study the effects of exogenously added anandamide on transient norepinephrine (NE)-induced contractions in mesenteric beds isolated from adult male Sprague-Dawley rats 6 h after the i.p. administration of 5 mg kg(-1) lipopolysaccharide (LPS). LPS treatment induced a 3-fold increase in total nitric-oxide synthase (NOS) activity without modifying either the systolic blood pressure or the vascular reactivity to NE of the isolated mesenteric bed. The endocannabinoid anandamide (0.01-10 microM) caused concentration-dependent reductions of the contractile responses to NE in the isolated mesenteric bed. This effect was significantly potentiated after LPS treatment compared with the controls. Anandamide-induced reductions of the contractile responses to NE in mesenteric beds isolated from LPS-treated rats were unmodified by endothelium removal but significantly diminished by either the anandamide amidase inhibitor phenylmethylsulfonyl fluoride (200 microM) or the vanilloid receptor antagonist capsazepine (1 microM). The vanilloid receptor agonist capsaicin (0.01-100 nM) also caused concentration-dependent relaxations that were potentiated in mesenteric beds from LPS-treated rats. Nevertheless, they were unmodified by 1 microM capsazepine. It is concluded that the potentiation of anandamide relaxations after LPS treatment, which are evident at early stages of endotoxic shock, could involve the participation of an anandamide metabolite and might be mediated, at least in part, through a vanilloid receptor.     

Bench-to-bedside review: Inhaled nitric oxide therapy in adults

Nitric oxide (NO) is an endogenous mediator of vascular tone and host defence. Inhaled nitric oxide (iNO) results in preferential pulmonary vasodilatation and lowers pulmonary vascular resistance. The route of administration delivers NO selectively to ventilated lung units so that its effect augments that of hypoxic pulmonary vasoconstriction and improves oxygenation. This 'Bench-to-bedside' review focuses on the mechanisms of action of iNO and its clinical applications, with emphasis on acute lung injury and the acute respiratory distress syndrome. Developments in our understanding of the cellular and molecular actions of NO may help to explain the hitherto disappointing results of randomised controlled trials of iNO.