BACKGROUNDAcute lung injury/acute respiratory distress syndrome presents with not only local inflammation, but also pulmonary coagulopathy which is characterized by an alveolar procoagulant response, anticoagulant inhibition, fibrinolytic supression and fibrin deposition. We thus had hypothesized that if aerosolized unfractionated heparin was inhaled into alveolar spaces, it could block the procoagulant tendency, lessen depletion of coagulation factors, and even influence the inflammatory response. We also assessed the effects of different administration regimens of heparin.

METHODSMale Wistar rats were given inhaled heparin starting 30 minutes before (prophylactic heparin) or 2 hours after (therapeutic heparin) intravenous lipopolysaccharide (LPS) was administered at 6-hour intervals; control groups received inhaled normal saline with or without being exposed to LPS. Thrombin-antithrombin complexes, activated protein C, tissue type and urokinase type plasminogen activators (t-PA/u-PA), plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor-α, interleukin-6 in bronchoalveolar lavage, and lung tissue myeloperoxidase activity, and histology score were measured at three time-points. PAI-1/(t-PA + u-PA) was calculated based on the before-mentioned parameters. Statistical analysis was made using one-way analysis of variance (ANOVA) with post hoc test or Student's t test in the case of heterogeneity of variance.

RESULTSAn alveolar procoagulant reaction, depressed fibrinolysis, and inflammatory response occurred in endotoxemia-induced lung injury. Local prophylactic application of heparin attenuated coagulation and early inflammation, promoted fibrinolysis, and reduced the histology score. Therapeutic application of heparin had similar, but weaker effects.

CONCLUSIONSIntrapulmonary application of unfractionated heparin by inhalation might inhibit alveolar procoagulant reaction and the early inflammatory response, promote fibrinolysis, and alleviate pulmonary pathology in endotoxemia-induced lung injury rats. Administration of heparin before LPS challenge was more efficacious.

Acute Lung Injury ; blood ; drug therapy ; Administration, Inhalation ; Animals ; Blood Coagulation ; drug effects ; Endotoxemia ; complications ; Fibrinolysis ; drug effects ; Heparin ; administration & dosage ; Inflammation ; drug therapy ; Lung ; pathology ; Male ; Rats ; Rats, Wistar

This study is to investigate the effects of sophoridine on NF-kappaB signaling pathway in kidney tissue of endotoxemia mice and the mechanism involved. BALB/c mice were challenged with lipopolysaccharide (LPS) caudal vein injection, then sophoridine was administered by intraperitoneal injection. Totally 50 mice were divided into 5 groups: control group, LPS model group, sophoridine treatment 12 mg x kg(-1) group, 6 mg x kg(-1) group and 3 mg x kg(-1) group. All animals were sacrificed at 6 hours after treatment. Kidney and blood samples were harvested. IKKbeta mRNA and TNF-alpha mRNA expression of renal tissue was measured by the reverse transcription polymerase chain reaction (RT-PCR), and phosphorylation IKKbeta protein (pIKKbeta) was detected by immunohistochemistry. NF-kappaB P65 protein expression and distribution of renal tissue were observed by Western blotting and immunofluorescence laser confocal microscopy. Serum TNF-alpha level was detected by radioimmunoassay. The results showed that the sophoridine significantly reduced the expression of IKKbeta mRNA and pIKKbeta protein, and inhibited the expression of NF-kappaB P65 protein and decreased the entry nuclear rate of NF-kappaB P65 in the renal tissue of endotoxemia mice. Thereby the renal TNF-alpha mRNA expression and serum TNF-alpha level were significantly reduced. These results suggest that sophoridine could inhibit inflammatory reaction induced by LPS through inhibiting activation of NF-kappaB signaling pathway.
Alkaloids ; pharmacology ; Animals ; Antitoxins ; pharmacology ; Endotoxemia ; blood ; chemically induced ; genetics ; metabolism ; Female ; I-kappa B Kinase ; genetics ; metabolism ; Kidney ; metabolism ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Phosphorylation ; Quinolizines ; pharmacology ; RNA, Messenger ; metabolism ; Random Allocation ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; genetics ; metabolism

OBJECTIVEGram-negative bacteria-induced multiple organ failure/dysfunction syndrome (MOF/MODS) is one of the leading causes of death through the world. The member of immunoglobulin family programmed death-1 (PD-1) is a negative immune regulator. This study investigated the protective effect of PD-1 as well as the underlying mechanism in LPS-induced endotoxemia.

METHODSTen PD-1(+/+) and ten PD-1 knockout (PD-1(-/-)) mice were injected peritoneally with LPS (10 mg/kg), and the survival was observed within 72 hrs after LPS injection. The other 40 PD-1(+/+) and 40 PD-1(-/-) mice were injected peritoneally with LPS (5 mg/kg). Blood samples were collected before injection and 1.5, 3 and 6 hrs after LPS injection (n=10 each time point). Serum levels of various inflammatory mediators were measured using ELISA.

RESULTSThe survival rate in PD-1(-/-) mice was noticeably lower than that in PD-1(+/+) mice after 10 mg/kg LPS injection. Serum levels of inflammatory mediators TNF-α, IL-1β, IL-12 and IL-17 in PD-1/mice were higher than those in PD-1(+/+) mice after 5 mg/kg LPS injection.

CONCLUSIONSPD-1 can protect mice from LPS-induced endotoxemia probably through its regulation on inflammatory mediator production.

Animals ; Antigens, Surface ; physiology ; Apoptosis Regulatory Proteins ; physiology ; Endotoxemia ; prevention & control ; Female ; Interleukin-12 ; blood ; Interleukin-17 ; blood ; Interleukin-1beta ; blood ; Lipopolysaccharides ; toxicity ; Mice ; Programmed Cell Death 1 Receptor ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the effect of glutamine on intestinal barrier function by examining the changes of plasma D-lactic levels and diamine oxidase (DAO) levels in plasma and intestinal tissue after glutamine intervention in young rats with endotoxemia.

METHODSEighty 18-day-old rats were randomly divided into endotoxemia and glutamine intervention groups (n=40 each). Endotoxemia was induced by lipopolysaccharide (LPS) injection. Plasma and small intestine homogenate were collected 1.5, 6, 24 and 72 hrs and 7 days after LPS injection. The glutamine intervention group was immediately administered with oral glutamine (2 g/kg) after LPS injection. Afterwards, glutamine was administered once daily. Plasma D-lactic and DAO levels and intestinal DAO levels were measured.

RESULTSPlasma DAO activity in the glutamine intervention group was significantly lower than that in the endotoxemia group 6 and 72 hrs after LPS injection (P<0.05). In contrast, the intestinal DAO activity in the glutamine intervention group was significantly higher than that in the endotoxemia group 6, 24 and 72 hrs and 7 days after LPS injection (P<0.05 or 0.01). Plasma D-lactic levels in the glutamine intervention group were significantly lower than those in the endotoxemia group 6, 24 and 72 hrs and 7 days after LPS injection (P<0.01).

CONCLUSIONSGlutamine may reduce the permeability of intestinal mucosa, and thus provides protective effects on intestinal barrier function in rats with endotoxemia.

Administration, Oral ; Amine Oxidase (Copper-Containing) ; metabolism ; Animals ; Behavior, Animal ; Endotoxemia ; metabolism ; Female ; Glutamine ; administration & dosage ; pharmacology ; Intestines ; drug effects ; metabolism ; Lactic Acid ; blood ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study hepatic NF-κB level following endotoxemic liver injury, and its relationship with hepatic TNF-α and IL-6 levels in young rats.

METHODSForty 18-day-old rats were randomly assigned to a normal control and an endotoxemia group. Endotoxemia was induced by lipopolysaccharide injection (LPS, 5 mg/kg). The endotoxemia group was subdivided into four groups sampled at 2, 6, 12 and 24 hrs after LPS injection (n=8 each). Pathological changes in liver cells were observed under a light microscope. TNF-α and IL-6 levels in liver tissue homogenates were measured using ELISA. Reitman-Frankel was used to measure serum ALT concentrations. NF-κB activation level in liver tissues was detected by immunohistochemistry.

RESULTSLiver tissue injury was the most obvious 6 hrs after LPS injection under the light microscope, and the damage rating of liver tissues was significantly higher in the endotoxemia group than that in the normal control group at all time points (P<0.05). ALT levels in the endotoxemia group were significantly higher than those in the normal control group 6, 12 and 24 hrs after LPS injection (P<0.05). NF-κB p65 protein expression in liver cells (percentage of nuclear positive cells) in the endotoxemia groups was significantly higher than that in the normal control group (P<0.05). TNF-α and IL-6 levels in liver tissue homogenates in the endotoxemia groups were significantly higher than those in the normal control group 6 and 12 hrs after LPS injection (P<0.05).

CONCLUSIONSEndotoxemia can cause liver injury, resulting in liver cell damage and changes in liver function. NF-κB activation is involved in endotoxemic liver injury which may be mediated by inflammatory cytokines TNF-α and IL-6 synthesis.

Alanine Transaminase ; blood ; Animals ; Behavior, Animal ; Endotoxemia ; complications ; Female ; Interleukin-6 ; analysis ; Liver ; chemistry ; pathology ; Liver Diseases ; etiology ; metabolism ; Male ; NF-kappa B ; analysis ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; analysis

Alanine Transaminase ; blood ; Animals ; Bilirubin ; blood ; Disease Models, Animal ; Endotoxemia ; etiology ; Endotoxins ; blood ; Gastrointestinal Hormones ; blood ; Gastrointestinal Motility ; physiology ; Intestine, Small ; physiopathology ; Liver ; pathology ; Liver Failure, Acute ; blood ; etiology ; physiopathology ; Liver Function Tests ; Rats ; Thioacetamide ; administration & dosage

Atelectasis can impair arterial oxygenation and decrease lung compliance. However, the effects of atelectasis on endotoxemic lungs during ventilation have not been well studied. We hypothesized that ventilation at low volumes below functional residual capacity (FRC) would accentuate lung injury in lipopolysaccharide (LPS)-pretreated rats. LPS-pretreated rats were ventilated with room air at 85 breaths/min for 2 hr at a tidal volume of 10 mL/kg with or without thoracotomy. Positive end-expiratory pressure (PEEP) was applied to restore FRC in the thoracotomy group. While LPS or thoracotomy alone did not cause significant injury, the combination of endotoxemia and thoracotomy caused significant hypoxemia and hypercapnia. The injury was observed along with a marked accumulation of inflammatory cells in the interstitium of the lungs, predominantly comprising neutrophils and mononuclear cells. Immunohistochemistry showed increased inducible nitric oxide synthase (iNOS) expression in mononuclear cells accumulated in the interstitium in the injury group. Pretreatment with PEEP or an iNOS inhibitor (1400 W) attenuated hypoxemia, hypercapnia, and the accumulation of inflammatory cells in the lung. In conclusion, the data suggest that atelectasis induced by thoracotomy causes lung injury during mechanical ventilation in endotoxemic rats through iNOS expression.
Animals ; Blood Pressure ; Carbon Dioxide/blood ; Cardiac Output ; Combined Modality Therapy ; Endotoxemia/*complications/immunology/pathology ; Functional Residual Capacity ; Immunohistochemistry ; Leukocytes, Mononuclear/pathology ; Lipopolysaccharides/pharmacology ; Lung/enzymology/pathology/physiopathology ; Lung Compliance ; Lung Volume Measurements ; Male ; Neutrophils/pathology ; Nitric Oxide Synthase Type II/metabolism ; Oxygen/blood ; Positive-Pressure Respiration/*adverse effects ; Pulmonary Atelectasis/*etiology/pathology/*therapy ; Rats ; Rats, Sprague-Dawley ; Thoracotomy/*adverse effects

OBJECTIVETo observe the effects of stimulation of intact vagus nerve (IVNS) on systemic inflammatory response in rats.

METHODSThe model of systemic inflammatory response was reproduced by lipopolysaccharide (LPS). One hundred Sprague Dawley rats were randomly divided into A group (with intravenous injection of LPS), B group (with stimulation of efferent nerve trunk of vagus nerve after intravenous injection of lipopolysaccharide and vagotomy), C group (with stimulation of intact vagus nerve after intravenous injection of LPS), D group (with vagus nerve stimulation after injection of equivalent isotonic saline), E group (with intravenous injection of equivalent isotonic saline), with 20 rats in each group. Five rats of each group were used to determine mean aortic pressure (MAP) before injection and l0 min, 0.5 h, 1 h, 2 h, 4 h and 6 h after injection. The serum levels of tumor necrosis factor (TNF) alpha and interleukin (IL) 10 were determined with enzyme linked immunosorbent assay before injection and 2, 4 and 6 hour after injection.

RESULTSThe level of MAP rose in A, B, C groups at 10 min after injection, especially in A group (134.40 +/- 7.3 mm Hg, l mm Hg = 0.033 kPa, P < 0.05), but it dropped in above groups at 30 min after injection. The level of MAP in A group was obviously lower than that in B, C groups during 10 min -6 h after injection. The serum level of TNF-alpha in A group was significantly higher than that in B, C groups at 2, 4, 6 hours after injection (P < 0.05). Compared with that in C group, the serum level of IL-10 in A, B groups lowered markedly at 4, 6 hours after injection (P < 0.05).

CONCLUSIONIVNS can stabilize hemodynamics and exert have anti-inflammatory effects at early stage of systemic inflammatory response.

Animals ; Blood Pressure ; Endotoxemia ; blood ; therapy ; Interleukin-10 ; blood ; Lipopolysaccharides ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood ; Vagus Nerve Stimulation

OBJECTIVETo investigate the changes in the plasma levels of endotoxin in severe burn patients during administration of antibiotics.

METHODSFifty severe burn patients with burn area larger than 30% TBSA were enrolled in the study, and they were respectively treated with Netilmicin (A group), Cefoperazone (B group), Ceftazidime (C group) and Imipenem/Cilastatin (D group). Venous blood samples were harvested for determination of endotoxins levels before treatment and 1, 2, 3, 5, 7 post-treatment day (PTD).

RESULTSThe plasma levels of endotoxin were elevated in different degrees in A, B and C groups. The plasma levels of endotoxin in B group were higher on 1, 2 PTD than on 3, 5, 7 PTD, and they were also higher than that in D group (P < 0.05). The plasma levels of endotoxin in C group reached the peak on 5 PTD [(0.398 +/- 0.172) EU/mL], which were higher than that before treatment [(0.251 +/- 0.142) EU/mL, P < 0.05] and other groups (P < 0.05). The plasma levels of endotoxin in D group were lower on 1, 2 PTD than that before treatment (P < 0.05).

CONCLUSIONDifferent amounts of endotoxins can be released after treatment with antibiotics in severe burn patients. Attention should be paid to the effect of antibiotics on the levels of endotoxin in practice.

Adolescent ; Adult ; Anti-Bacterial Agents ; therapeutic use ; Burns ; blood ; drug therapy ; Endotoxemia ; etiology ; Endotoxins ; blood ; Female ; Humans ; Male ; Middle Aged ; Plasma ; Young Adult

In light of the anti-inflammatory properties of histone deacetylase (HDAC) inhibitors, such as suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA), we examined a new HDAC inhibitor KBH-A42 for its anti-inflammatory activities. KBH-A42 showed noteworthy anti-inflammatory properties in vitro via suppression of the production of TNF-alpha, a proinflammatory cytokine, and nitric oxide (NO), a proinflammatory effector molecule, in LPS-stimulated RAW264.7 cells and peritoneal macrophages. It also inhibited TNF-alpha production in vivo as demonstrated in a LPS-induced mouse endotoxemia model. The levels of TNF-alpha, IL-1beta, IL-6 and iNOS mRNAs determined by RT-PCR propose that the inhibition of these pro-inflammatory mediators by KBH-A42 resulted from inhibiting expression of these genes. However, the EMSA study to see the effect of KBH-A42 on the binding of NF-kappaB, a transcription factor, to a specific DNA sequence showed that the binding of NF-kappaB to DNA was not changed regardless of increasing the concentration of KBH-A42 in the presence and absence of LPS stimulation. Interestingly, DNA binding of another transcription factor AP-1 dose-dependently increased by KBH-A42. KBH-A42 differentially regulated the phosphorylation of MAP kinases. While the phosphprylation of ERK1/2 and SAPK/JNK was not affected by KBH-A42, the phosphorylation of p38 decreased by KBH-A42. These results showed that KBH-A42 inhibits production of proinflammatory cytokines in macrophages by decreasing their mRNA levels, and p38 kinase is involved in the KBH-A42-mediated inhibition.
Animals ; Blotting, Western ; Cell Line ; Cell Survival/drug effects ; Cytokines/blood/genetics/*metabolism ; Electrophoretic Mobility Shift Assay ; Endotoxemia/blood/metabolism/pathology ; Enzyme Inhibitors/chemistry/*pharmacology ; Histone Deacetylases/*antagonists & inhibitors ; Hydroxamic Acids/chemistry/*pharmacology ; Interleukin-1beta/genetics/metabolism ; Interleukin-6/genetics/metabolism ; Macrophages/cytology/*drug effects/metabolism ; Mice ; Mitogen-Activated Protein Kinase 1/metabolism ; Mitogen-Activated Protein Kinase 3/metabolism ; Mitogen-Activated Protein Kinases/metabolism ; Molecular Structure ; NF-kappa B/metabolism ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type II/genetics/metabolism ; Phosphorylation/drug effects ; Piperidones/chemistry/*pharmacology ; Protein Binding/drug effects ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor AP-1/metabolism ; Tumor Necrosis Factor-alpha/blood/genetics/metabolism