8-epi-prostaglandin-f2alpha and Sepsis

Sepsis commonly progresses to acute lung injury and is associated with high morbidity and mortality. Septic acute lung injury is characterized by severe oxidative stress response, remained refractory to present therapies, and new therapies need to be developed to improve further clinical outcomes. We determined the effect of betulinic acid (BA) on oxidative lung injury in mice using cecal ligation and puncture (CLP) model.. Five groups of mice (six in each group) received three pretreatments at 24-h interval before surgery. Surgery was done 1 h after last dosing. Sham and CLP control group mice received vehicle. BA was administered to other three groups of mice at 3, 10, and 30 mg/kg dose. Lung and plasma samples were collected for analysis by sacrificing the mice at 18 h of surgery.. Compared with sham, CLP significantly increased total protein, nitrite, malondialdehyde, isoprostane, superoxide, protein carbonyl, oxidative stress index, inducible nitric oxide synthase protein, and histopathologic changes and reduced the superoxide dismutase, catalase activity, and total thiol levels in lungs and plasma, which were restored by BA pretreatment.. BA pretreatment decreased the levels of oxidants, increased the levels of antioxidants in lungs and plasma thereby reducing the oxidative lung injury in CLP mice. Additionally, BA was found to scavenge the superoxide and nitric oxide radical in vitro. Thus, BA is suggested to be effective in treatment of oxidative lung injury in sepsis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Betulinic Acid; Dinoprost; Drug Evaluation, Preclinical; Lung; Lung Injury; Male; Malondialdehyde; Mice; Nitric Oxide; Nitric Oxide Synthase Type II; Nitrites; Oxidative Stress; Pentacyclic Triterpenes; Protein Carbonylation; Sepsis; Superoxides; Triterpenes

To investigate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) in the protective effects of hydrogen against cerebral dysfunction in a mouse model of sepsis.. Male ICR mice were randomly divided into sham operation group, hydrogen control group, sepsis group and hydrogen treatment group, with 20 in each group. Sepsis model was reproduced by cecal ligation and puncture (CLP). 2% hydrogen inhalation was given for 1 hour at 1 hour and 6 hours after operation in hydrogen treatment group. The brain tissues were obtained at 24 hours after operation. The histopathologic changes and neuron apoptosis in the hippocampus were observed under the microscope. The expressions of nucleus and total Nrf2 in hippocampus were detected by Western Blot. The activities of superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and 8-iso-prostaglandin F2α (8-iso-PGF2α) in hippocampus were also detected. The changes of learning and memory abilities were observed by Morris water maze test at day 4 to 9 after operation.. Compared with the sham operation and hydrogen control groups, in the sepsis group, the number of normal pyramidal neurons in the hippocampal CA1 region was markedly reduced, the apoptotic index was marked increased, the expressions of nucleus and total Nrf2 were partly increased, the activities of SOD and CAT in the hippocampus were significantly decreased, and the levels of MDA and 8-iso-PGF2α were markedly increased, the escape latency at day 4 to 8 after operation was significantly extended, and there was no difference in swimming speed, the percentage of time in the target quadrant and the times of the platform crossing were significantly decreased on probe day. Compared with the sepsis group, in the hydrogen treatment group, the number of normal pyramidal neurons in the hippocampal CA1 region was markedly increased (67.33 ± 6.89 vs. 42.33 ± 6.02, P<0.01), the apoptotic index was dramatically reduced [(30.00 ± 4.77)% vs. (80.50 ± 6.99)%, P<0.01], the expressions of nucleus and total Nrf2 were significantly increased [nucleus Nrf2 (A value): 5.07 ± 0.35 vs. 3.04 ± 0.34, total Nrf2 (A value): 4.24 ± 0.58 vs. 2.91 ± 0.37, both P<0.01], the activities of SOD and CAT in the hippocampus were significantly increased [SOD (U/mg): 120.96 ± 13.44 vs. 81.16 ± 12.28, CAT (U/mg): 9.11 ± 1.28 vs. 5.64 ± 1.88, both P<0.01], and the levels of MDA and 8-iso-PGF2α were markedly reduced [MDA (nmol/mg): 16.12 ± 1.49 vs. 27.64 ± 1.87, 8-iso-PGF2α (pg/mg): 183.43 ± 13.07 vs. 864.07 ± 49.92, both P<0.01], the escape latency at day 5 to 8 after operation was significantly shortened, and there was no difference in swimming speed, the percentage of time in the target quadrant [(37.06 ± 1.16)% vs. (24.42 ± 1.82)%, P<0.01] and the times of the platform crossing (7.13 ± 0.98 vs. 4.88 ± 0.99, P<0.01) were significantly increased on probe day. There was no statistical difference in above indexes between sham operation group and hydrogen control group.. Hydrogen inhalation can ameliorate pathological injury in brain and impairment of learning and memory abilities of septic mice, which may be associated with the up-regulation of Nrf2, the increase of antioxidant enzymes activities and the decrease of oxidative products.

Topics: Animals; Brain; Dinoprost; Disease Models, Animal; Hydrogen; Male; Malondialdehyde; Mice; Mice, Inbred ICR; NF-E2-Related Factor 2; Sepsis; Superoxide Dismutase

Sepsis is the systemic response of an organism against microorganisms and toxins. Lithium is a therapeutic agent used for bipolar disorder and neurodegenerative disease, and it exerts pleiotropic effects on various cellular processes. The present study aimed to determine the effect of lithium on cecal ligation and puncture (CLP)-induced tissue injury in the lungs, by inhibiting the pro-inflammatory cytokine response, and the generation of reactive oxygen species (ROS) triggered by polymicrobial sepsis. Five groups of 20 rats each were used: 1) sham-operated control group; 2) CLP group; 3) 50mg/kg lithium-treated control healthy group; 4) 25 mg/kg lithium-treated CLP group; and 5) 50 mg/kg lithium-treated CLP group. A CLP polymicrobial sepsis model was applied to the rats. All rat groups were killed 16 h later, and lung and blood samples were analyzed histopathologically and biochemically. The 25 and 50 mg/kg of lithium decreased the level of interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and the tumor necrosis factor-α (TNF-α) in the serum, and the 8-iso-prostaglandin F2α (8-ISO) level in lung tissue. The lithium also increased the activity of superoxide dismutase (SOD) and the total levels of glutathione (GSH) in the lung tissues of rats. The histopathological scores and examinations were in accordance with the biochemical results, and revealed significant differences in the inflammation scores between the sepsis group and the other groups. The CLP+lithium 50mg/kg group had the lowest inflammation score among the CLP groups. Our results indicated that the therapeutic administration of lithium prevented oxidative stress changes and cytokine changes, and also protected vital tissues.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Cytokines; Dinoprost; Disease Models, Animal; Glutathione; Lithium Carbonate; Male; Rats; Rats, Wistar; Sepsis; Superoxide Dismutase

Sepsis is a serious medical condition that is characterized by a whole-body inflammatory state and the presence of a known or suspected infection. Amiodarone is a class III antiarrhythmic agent, a multichannel blocker (Ca++, Na+, and K+), and a noncompetitive α- and β-adrenergic blocker in cardiac cells. The present study aimed to determine whether amiodarone was protective against experimentally induced cecal ligation and puncture sepsis in rat lung tissue. The relationship between its probable protective effect and antioxidant/anticytokine action biochemically and histopathologically was also examined. Five groups of rats were used, each composed of 20 rats: (1) the sham-operated control group; (2) the CLP group; (3) the 25-mg/kg amiodarone-treated control healthy group; (4) the 50-mg/kg amiodarone-treated CLP group; and (5) the 50-mg/kg amiodarone-treated CLP group. A CLP polymicrobial sepsis model was applied to the rats. All groups were sacrificed 16 h later, and lung and blood samples were analyzed histopathologically and biochemically. Twenty-five and 50 mg/kg amiodarone decreased the level of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α in serum and 8-iso-prostaglandin F2α level in lung tissue. They increased the activities of superoxide dismutase and levels of total glutathione in lung tissues of rats. Histopathological scores and examinations were in accordance with the biochemical results. Histopathological analysis revealed significant differences in inflammation scores between the sepsis group and the other groups. The CLP + amiodarone 50 mg/kg group had the lowest inflammation score among CLP groups. Our results indicate that administration of amiodarone prevented oxidative stress and cytokine action and protected lung tissue during sepsis cascade.

Topics: Amiodarone; Animals; Cecum; Cytokines; Dinoprost; Glutathione; Ligation; Lung; Male; Oxidative Stress; Protective Agents; Punctures; Rats; Rats, Wistar; Sepsis; Superoxide Dismutase

Sepsis is the most common cause of death in intensive care units. Some studies have found that hyperoxia may be beneficial to sepsis. However, the clinical use of hyperoxia is hindered by concerns that it could exacerbate organ injury by increasing free radical formation. Recently, it has been suggested that molecular hydrogen (H2) at low concentration can exert a therapeutic antioxidant activity and effectively protect against sepsis by reducing oxidative stress. Therefore, we hypothesized that combination therapy with H2 and hyperoxia might afford more potent therapeutic strategies for sepsis. In the present study, we found that inhalation of H2 (2%) or hyperoxia (98%) alone improved the 14-day survival rate of septic mice with moderate cecal ligation and puncture (CLP) from 40% to 80% or 70%, respectively. However, combination therapy with H2 and hyperoxia could increase the 14-day survival rate of moderate CLP mice to 100% and improve the 7-day survival rate of severe CLP mice from 0% to 70%. Moreover, moderate CLP mice showed significant organ damage characterized by the increases in lung myeloperoxidase activity, lung wet-to-dry weight ratio, protein concentration in bronchoalveolar lavage, serum biochemical parameters (alanine aminotransferase, aspartate aminotransferase, creatinine, and blood urea nitrogen), and organ histopathological scores (lung, liver, and kidney), as well as the decrease in PaO2/FIO2 ratio at 24 h, which was attenuated by either H2 or hyperoxia alone. However, combination therapy with H2 and hyperoxia had a more beneficial effect against lung, liver, and kidney damage of moderate or severe CLP mice. Furthermore, we found that the beneficial effect of this combination therapy was associated with the decreased levels of oxidative product (8-iso-prostaglandin F2α), increased activities of antioxidant enzymes (superoxide dismutase and catalase) and anti-inflammatory cytokine (interleukin 10), and reduced levels of proinflammatory cytokines (high-mobility group box 1 and tumor necrosis factor α) in serum and tissues. Therefore, combination therapy with H2 and hyperoxia provides enhanced therapeutic efficacy via both antioxidant and anti-inflammatory mechanisms and might be potentially a clinically feasible approach for sepsis.

Topics: Alanine Transaminase; Animals; Catalase; Coinfection; Cytokines; Dinoprost; Disease Models, Animal; Glutamyl Aminopeptidase; Hydrogen; Hyperoxia; Inflammation Mediators; Kidney; Liver; Lung; Male; Mice; Peroxidase; Sepsis; Superoxide Dismutase

Pulmonary oxidant stress is an important pathophysiologic feature of acute lung injury. It is unclear whether nitric oxide contributes to this oxidant stress. Thus, we examined the role of inducible nitric oxide synthase (iNOS) in pulmonary oxidant stress in murine sepsis and the differential contribution of different cellular sources of iNOS.. Randomized, controlled animal study.. Research laboratory of an academic institution.. Male iNOS+/+, iNOS-/- C57Bl/6 mice, and bone-marrow transplanted iNOS chimeric mice: +to- (wild-type iNOS+/+ donor bone-marrow transplanted into iNOS-/- recipient mice) and the reciprocal -to+ chimeras.. Animals were randomized to sepsis (n = 264), induced by cecal ligation and perforation, vs. naive groups (n = 138).. In septic iNOS-/- vs. wild-type iNOS+/+ mice, sepsis-induced pulmonary oxidant stress (33 +/- 11 [mean +/- sem] vs. 365 +/- 48 pg 8-isoprostane/mg protein, p < .01) and nitrosative stress (0.0 +/- 0.0 vs. 0.9 +/- 0.4 micromol 3-nitrotyrosine/mmol para-tyrosine, p < .05) were abolished, despite similar septic increases in pulmonary myeloperoxidase activity in both (86 +/- 20 vs. 83 +/- 12 mU/mg protein, p = .78). In +to- iNOS chimeric mice (iNOS localized only to donor bone-marrow-derived inflammatory cells), cecal ligation and perforation resulted in significant pulmonary oxidant stress (368 +/- 81 pg 8-isoprostane/mg protein) and nitrosative stress (0.6 +/- 0.2 micromol 3-nitrotyrosine/mmol para-tyrosine), similar in degree to septic wild-type mice. In contrast, pulmonary oxidant and nitrosative stresses were absent in septic -to+ iNOS chimeras (iNOS localized only to recipient parenchymal cells), similar to iNOS-/- mice.. In murine sepsis-induced acute lung injury, pulmonary oxidant stress is completely iNOS dependent and is associated with tyrosine nitration. Moreover, pulmonary oxidant stress and nitrosative stress were uniquely dependent on the presence of iNOS in inflammatory cells (e.g., macrophages and neutrophils), with no apparent contribution of iNOS in pulmonary parenchymal cells. iNOS inhibition targeted specifically to inflammatory cells may be an effective therapeutic approach in sepsis and acute lung injury.

Topics: Analysis of Variance; Animals; Bone Marrow Transplantation; Chimera; Dinoprost; Lung; Macrophages, Alveolar; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Oxidative Stress; Random Allocation; Respiratory Distress Syndrome; Sepsis; Tyrosine

We have recently demonstrated that selective inducible nitric oxide (NO) synthase (iNOS) inhibition with 1400W attenuated the hemodynamic and metabolic alterations affiliated with hyperdynamic porcine endotoxemia. In contrast to endotoxemia, limited evidence is available to document a relationship between NO and organ dysfunction in large animal bacteremic models. Therefore, using the same experimental setup, we investigated the role of selective iNOS blockade in porcine bacteremia induced and maintained for 24 h with a continuous infusion of live Pseudomonas aeruginosa. After 12 h of sepsis, animals received either vehicle (Control, n = 8) or continuous infusion of selective iNOS inhibitor, L-N6-(1-iminoethyl)-lysine (L-NIL; n = 8). Measurements were performed before, and 12, 18, and 24 h after P. aeruginosa infusion. L-NIL inhibited sepsis-induced increase in plasma nitrate/nitrite concentrations and prevented hypotension without affecting cardiac output. Despite comparable hepatosplanchnic macrocirculation, L-NIL blunted the progressive deterioration in ileal mucosal microcirculation and prevented mucosal acidosis. L-NIL largely attenuated mesenteric and hepatic venous acidosis, significantly improved P. aeruginosa-induced impairment of hepatosplanchnic redox state, and mitigated the decline in liver lactate clearance. Furthermore, the administration of L-NIL reduced the hepatocellular injury and prevented the development of renal dysfunction. Finally, treatment with L-NIL significantly attenuated the formation of 8-isoprostane concentrations, a direct marker of lipid peroxidation. Thus, selective iNOS inhibition with L-NIL prevented live bacteria from causing key features of metabolic derangements in porcine hyperdynamic sepsis. Underlying mechanisms probably include reduced oxidative stress with improved microcirculatory perfusion and restoration of cellular respiration.

Topics: Animals; Arteries; Bacteremia; Dinoprost; Endotoxemia; Intestinal Mucosa; Kidney; Lactates; Liver; Lysine; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Oxidative Stress; Oxygen; Pseudomonas aeruginosa; Pyruvates; Sepsis; Swine; Time Factors

Sepsis is known to be associated with oxidative stress. Novel markers of oxidative stress are now believed to be F2-isoprostanes which are produced in situ in phospholipids and subsequently released into circulation and excreted in the urine. This study, therefore, sought to investigate whether the excretion of the isoprostane, 8-iso-PGF(2 alpha), is elevated during sepsis. The excretion of 8-iso-PGF(2 alpha), in the 24 h urine of three patients was studied in the septic stage, during mobilisation and in the state of health by a radioimmunological method. Extrapolating the urinary excretion of 8-iso-PGF(2 alpha) over time showed an insignificant variation in the excretion values during 24 h. The amount of mean 24 h urinary 8-iso-PGF(2 alpha) was about similar in the septic stage and in the state of health but increased remarkably during mobilisation in two of the patients. We suggest that mobilisation of septic patients can be associated with an increase of oxidative stress which may stem from an increase in oxygen consumption and/or from a depletion of antioxidants leading to the enhanced formation of free radicals.

Topics: Dinoprost; F2-Isoprostanes; Health; Humans; Intensive Care Units; Oxidative Stress; Recovery of Function; Sepsis; Urinalysis

We hypothesized that endotoxin injection in rats would stimulate in vivo nuclear factor-kappa B (NF-kappa B) activation in lung tissue and that antioxidant treatment before endotoxin injection would attenuate endotoxin-induced NF-kappa B activation, chemokine gene expression, and neutrophilic lung inflammation. We studied NF-kappa B activation in rat lung tissue following a single i.p. injection of endotoxin (6 mg/kg). After in vivo endotoxin treatment, lung NF-kappa B activation peaked at 2 h and temporally correlated with the expression of cytokine-induced neutrophil chemoattractant mRNA in lung tissue. Treatment with the antioxidant N-acetylcysteine (NAC) 1 h before endotoxin resulted in decreased lung NF-kappa B activation in a dose-dependent manner (from 200-1000 mg/kg) and diminished cytokine-induced neutrophil chemoattractant mRNA expression in lung tissue. Treatment with NAC significantly suppressed endotoxin-induced neutrophilic alveolitis. The average total lung lavage neutrophil count was 5.5 x 10(6) with endotoxin treatment vs 0.9 x 10(6) with NAC treatment before endotoxin. The NF-kappa B pathway represents an attractive therapeutic target for strategies to control neutrophilic inflammation and lung injury.

Topics: Acetylcysteine; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Base Sequence; Chemokines, CXC; Chemotactic Factors; Chemotaxis, Leukocyte; Dinoprost; Disease Models, Animal; Drug Evaluation, Preclinical; Endotoxins; F2-Isoprostanes; Gene Expression Regulation; Glutathione; Growth Substances; Inflammation; Intercellular Signaling Peptides and Proteins; Leukocyte Count; Lung; Lung Diseases; Male; Molecular Sequence Data; Neutrophils; NF-kappa B; Rats; Rats, Sprague-Dawley; Respiratory Distress Syndrome; RNA, Messenger; Sepsis